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More frequent among adults, phenocopies may be caused by somatic mutations or anti-cytokine autoantibodies, mimicking the phenotypes of primary immunodeficiencies. A fourteen-year-old girl was referred for a two-year history of weight loss and multiple recurrent abscesses, complicated recurrent pneumonia, pyelonephritis, osteomyelitis, and septic shock, without fever. She had started with nausea, hyporexia, and weight loss, then with abscesses in her hands, knee, ankle, and spleen. She also developed a rib fracture and left thoracic herpes zoster. The patient was cachectic, with normal vital signs, bilateral crackles on chest auscultation, tumefaction of the knee joint, and poorly healed wounds in hands and chest, oozing a yellowish fluid. Chest computed tomography revealed multiple bilateral bronchiectases. Laboratory workup reported chronic anemia, leukocytosis, neutrophilia, mild lymphopenia, thrombocytosis, pan-hypergammaglobulinemia, and elevated acute serum reactants. Lymphocyte subsets were low but present. Mycobacterium tuberculosis was detected via polymerase chain reaction in a bone biopsy specimen from ankle osteomyelitis. Whole-exome sequencing failed to identify a monogenic defect. Interleukin-12 was found markedly elevated in the serum of the patient. Phosphorylation of STAT4, induced by increasing doses of IL-12, was neutralized by patient serum, confirming the presence of anti-IL12 autoantibodies. IL-12 and IL-23 are crucial cytokines in the defense against intracellular microorganisms, the induction of interferon-gamma production by lymphocytes, and other inflammatory functions. Patients who develop neutralizing serum autoantibodies against IL12 manifest late in life with weight loss, multiple recurrent abscesses, poor wound healing, and fistulae. Treatment with anti-CD20 monoclonal antibodies was effective.

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IL-39 (Interleukin-39) is a heterodimeric cytokine composed of IL-23p19 and EBI3 (Epstein-Barr virus-induced gene 3) subunits. Despite the evidence that correlates the role of IL-39 in regulating inflammation, its expression in the intestinal microenvironment of IBD (inflammatory bowel disease) patients is still unknown. Thus, this work was focused on characterizing relative mRNA (messenger RNA) IL-39 expression and intestinal synthesis in IBD patients. This study includes 37 patients diagnosed with ulcerative colitis (UC), 15 with Chron's disease (CD), and 22 controls. Gene expression of IL-39 subunits (IL-23p19/EBI3) was measured by RT-PCR (real time polymerase chain reaction). Intestinal synthesis was evaluated by immunohistochemistry and serum levels by ELISA. Statistical analysis was done using Prism GraphPad V6. Relative mRNA IL-39 expression was increased in patients with active UC and active CD compared to the remission UC, remission CD, and control group. High levels of relative mRNA expression of IL-39 (IL-23p19 subunit) were associated with histological activity. IHQ analysis showed increased IL-39 production in mucosa, submucosa, muscular, and serosa layer of patients with active disease. IL-39 serum production was increased in patients with UC. IL-39 gene's upregulation was found in patients with active IBD and was associated with severe histological activity in UC. This is the first report regarding the role of IL-39 in patients with IBD. The findings suggest that IL-39 might play a role as an inflammatory mediator in active IBD and could be considered a new alternative in treating this condition.

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BACKGROUND: The aim of this feasibility study was to investigate the concentration level of CCL-20/MIP-3α, BAFF/BLyS, IL-23, RANKL, and Osteoprotegerin in the Peri-Implant Crevicular Fluid (PICF), from patients diagnosed with peri-implant mucositis and peri-implantitis, and to compare them with PICF from patients with healthy implants. METHODS: Participants with at least one dental implant with healthy peri-implant tissues, peri-implant mucositis, or peri-implantitis were included. PICF was collected using paper strips from healthy and diseased peri-implant sites (n = 19). Biomarker levels were analyzed using a custom Multiplex ELISA Assay Kit. RESULTS: In comparison to peri-implant health, the peri-implant mucositis group showed an increased concentration of CCL-20 MIP-3α, BAFF/BLyS, IL-23, RANKL, and Osteoprotegerin. The peri-implantitis group had the lowest median concentration of Osteoprotegerin (1963 ng/mL); this group had a similar concentration of RANKL (640.84 ng/mL) when compared to the peri-implant health group. BAFF/BLyS (17.06 ng/mL) showed the highest concentration in the peri-implantitis group. CONCLUSIONS: This feasibility study suggests that IL-23 and RANKL may help to elucidate the pathogenesis during the conversion from peri-implant health to peri-implantitis. Further research is required in BAFF/BLyS for the early diagnosis of peri-implantitis.

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BACKGROUND: Juvenile systemic lupus erythematosus (jSLE) is known to be more severe and with a higher frequency of renal and central nervous system impairment when compared to systemic lupus erythematosus in adults. The study of immunological characteristics of jSLE patients might help to envisage better treatment strategies to reduce the burden of the disease. OBJECTIVE: To characterize peripheral lymphocytes, assessing activation markers, and PD-1 expression on T cells; to evaluate in vitro cytokine expression upon stimulation in jSLE patients and age-matched controls. METHODOLOGY: Eighteen jSLE patients on low disease activity and 25 matched healthy adolescents were evaluated for immune activation and PD-1 expression on peripheral blood lymphocytes by flow cytometry. Twenty-one cytokines were assessed by X-MAP technology after in vitro stimulation of peripheral blood with phytohemagglutinin. RESULTS: jSLE patients had lower numbers of CD4 T, CD8 T, B, and NK cells; higher central memory CD8 T cell percentages were noted in jSLE adolescents in comparison with controls (p = 0.014). B cells subsets showed a higher percentage of exhausted memory subset than controls (p = 0.014). The expression of PD-1 on CD4 T and CD8 T cells did not show relevant changes in jSLE adolescents. After stimulation of peripheral blood, cell supernatant of jSLE patients showed a trend to lower concentrations of IL-10 (p=0.080) and higher concentrations of IL-23 (p = 0.063) than controls. CONCLUSIONS: jSLE patients on low disease activity maintain lymphopenia of all subsets, with a B cell profile of exhaustion. Upon in vitro stimulation, peripheral blood cell supernatant showed a shift to IL-23, suggesting a role of inhibitors of this cytokine as another potential therapeutic target for those patients.

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BACKGROUND: Emerging evidence suggests that the interleukin (IL) 17/ IL-23 axis may play a role in the pathogenesis of leishmaniasis. Our aim was to investigate whether the IL-23R variant rs11805303 is a risk factor for the development of cutaneous leishmaniasis (CL) in Leishmania guyanensis-infected individuals. METHODS: We genotyped by polymerase chain reaction-restriction fragment length polymorphism the rs11805303 C/T in 828 patients with CL and 806 healthy individuals. Plasma tumor necrosis factor-α, IL-6, interferon-γ, IL-1ß, and IL-17 were measured with the Bioplex assay. RESULTS: The distribution of the genotypes differed between patients with CL and healthy controls with a common odds ratio of 1.78 (P = 2.2 × 10-11) for the disease-associated T allele. Leishmania guyanensis-infected individuals homozygous for the T allele show a 200% increased risk of progressing to disease development, with a 95% confidence interval ranging from 81% to 400% (P = 9.9 × 10-6) in comparison to individuals homozygous for the C allele. Males homozygous for the T allele have higher plasma levels of IL-17 compared with heterozygous or homozygous CC individuals. CONCLUSIONS: The present association of the IL-23R variant rs11805303 with the development of CL suggests that the IL-17/IL-23 axis may play an important role in the pathogenesis of CL.

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Rheumatoid arthritis (RA) is an autoimmune disease that causes joint destruction. Although its etiology remains unknown, citrullinated proteins have been considered as an auto-antigen able to trigger an inflammatory response in RA. Herein, we modified the classical antigen-induced arthritis (AIA) model by using citrullinated human plasma fibrinogen (hFIB) as an immunogen to investigate the mechanism of inflammation-driven joint damage by citrullinated hFIB in C57BL/6 mice. We found that hFIB-immunized mice showed high serum levels of anti-citrullinated peptides antibodies (ACPAs). Moreover, hFIB immunized mice showed increased mechanical hyperalgesia, massive leukocyte infiltration, high levels of inflammatory mediators, and progressive joint damage after the intra-articular challenge with citrullinated hFIB. Interestingly, hFIB-induced arthritis was dependent on IL-23/IL-17 immune axis-mediated inflammatory responses since leukocyte infiltration and mechanical hyperalgesia were abrogated in Il17ra-/- and Il23a-/- mice. Thus, we have characterized a novel model of experimental arthritis suitable to investigate the contribution of ACPAs and Th17 cell-mediated immune response in the pathogenesis of RA.

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Monoclonal antibodies or fusion proteins, defined as biological drugs, have modified the natural history of numerous immune-mediated disorders, allowing the development of therapies aimed at blocking the pathophysiological pathways of the disease, providing greater efficacy and safety than conventional treatment strategies. Virtually all therapeutic proteins elicit an immune response, producing anti-drug antibodies (ADAs) against hypervariable regions of immunoglobulins. Immunogenicity against biological drugs can alter their pharmacokinetic and pharmacodynamic properties, thereby reducing the efficacy of these drugs. In more severe cases, ADAs can neutralize the therapeutic effects of the drug or cause serious adverse effects, mainly hypersensitivity reactions. The prevalence of ADAs varies widely depending on the type of test used, occurrence of false-negative results, and non-specific binding to the drug, making it difficult to accurately assess their clinical impact. Concomitant use of immunosuppressors efficiently reduces the immunogenicity in a dose-dependent manner, either by decreasing the frequency of detectable ADAs or by delaying their appearance, thereby enhancing the effectiveness of biological therapies. Among the new therapeutic strategies for the management of psoriasis, biological agents have gained increasing importance in recent years as they interrupt key inflammation pathways involved in the physiopathology of the disease. Reports regarding ADA in new biologics are still scarce, but the most recent evidence tends to show little impact on the clinical response to the drug, even with prolonged treatment. It is therefore essential to standardize laboratory tests to determine the presence and titles of ADAs to establish their administration and management guidelines that allow the determination of the real clinical impact of these drugs.

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AIM: To determine the methylation pattern of TLR2 gene promoter and its association with the transcriptional regulation of periapical inflammatory and angiogenic responses in symptomatic and asymptomatic forms of apical periodontitis. METHODOLOGY: In this cross-sectional study, apical lesions were obtained from volunteers with asymptomatic apical periodontitis (AAP) (n = 17) and symptomatic apical periodontitis (SAP) (n = 17) scheduled for tooth extraction, and both total RNA and DNA were extracted. DNA was bisulfite-treated, a region of CpG island within the TLR2 gene was amplified by qPCR and the products were sequenced. Additionally, the mRNA expression of TLR2, TLR4, IL-6, IL-12, TNFalpha, IL-23, IL-10, TGFbeta, VEGFA and CDH5 was analysed by qPCR. The data were analysed with chi-square tests, Mann-Whitney or unpaired t-tests, and Spearman´s correlation; variable adjustments were performed using multiple linear regression (P < 0.05). RESULTS: TLR2 depicted a hypomethylated DNA profile at the CpG island in SAP when compared with AAP, along with upregulated expression of TLR2, with pro-inflammatory cytokines IL-6 and IL-23, and the angiogenesis marker CDH5 (P < 0.05). TLR2 methylation percentage negatively correlated with mRNA levels of IL-23 and CDH5 in apical periodontitis. Lower methylation frequencies of single CpG dinucleotides -8 and -10 localized in close proximity to nuclear factor κB (NFκB) binding within the TLR2 promoter were identified in SAP versus AAP (P < 0.05). Finally, unmethylated -10 and -8 single sites demonstrated up-regulation of IL-23, IL-10 and CDH5 transcripts compared to their methylated counterparts (P < 0.05). CONCLUSIONS: TLR2 gene promoter hypomethylation was linked to transcriptional activity of pro-inflammatory cytokines and angiogenic markers in exacerbated periapical inflammation. Moreover, unmethylated single sites in close proximity to NFκB binding were involved in active transcription of IL-23, IL-10 and CDH5.

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The Th17 profile immune response is influenced by the presence of cytokines such as IL-1, IL-6, TGF-ß, IL-17, and IL-23. We sought to characterize the Th17 profile in CNS samples from human rabies cases transmitted by dogs and examine its possible influence on disease pathogenesis. We observed a high expression of TGF-ß, followed by IL-23, IL-17 and IL-6, and a low expression of IL-1ß and IFN-γ. Those results suggest the participation of Th17 in rabies virus neuroinfection transmitted by dogs. IL-23 probably plays a role in maintaining the Th17 profile, but it can also interfere with the establishment of the Th1 profile and viral clearance.

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A tireoide é uma glândula endócrina que pode ser afetada por diversas lesões, que incluem doenças reativas, autoimunes e neoplásicas. Evidências mostram que a resposta imunológica, a partir da liberação de citocinas inflamatórias por células imunes e não imunes, desempenha um importante papel no desenvolvimento de diversas doenças. Dentre essas citocinas, destacam-se a IL-17 e a IL-23, as quais têm sido associadas à resposta Th17 e patogênese de diversas doenças tireoidianas. Dessa forma, esta pesquisa teve como objetivo avaliar a imunoexpressão das proteínas IL-17 e IL-23 em carcinomas papilíferos de tireoide (CPTs), adenomas foliculares (AFs), bócios coloides (BCs) e tireoidites de Hashimoto (THs) (isoladas e associadas a CPTs ou BCs), com o intuito de compreender melhor a atuação destas citocinas nestas entidades. A amostra foi composta por 30 casos de CPTs, 10 AFs, 15 BCs e 15 THs. A análise da imunoexpressão das proteínas nas células foliculares/tumorais foi realizada semiquantitativamente em toda extensão das lesões, as quais foram classificadas em três escores: 1 (≤ 33%), 2 (> 33% ­ 66%) e 3 (> 66%). Para a avaliação dos linfócitos imunopositivos, foram selecionados cinco campos com maior quantidade de linfócitos positivos, onde foi realizada a contagem. Os dados foram submetidos à análise estatística por meio dos testes de Kruskal-Wallis (KW), Mann-Whitney (U) e Spearman (r), com o nível de significância estabelecido em 5% (p < 0,05). Na análise da IL-17 entre as lesões, foi verificada maior imunoexpressão pelos linfócitos nas THs comparado aos AFs e BCs (p=0,041 e p=0,013). Na avaliação da IL-23 foi constatada uma maior imunomarcação das células foliculares/tumorais nas THs em relação aos CPTs (p=0,002), assim como, observou-se uma maior expressão desta citocina nos linfócitos das THs em comparação aos CPTs, AFs e BCs (p=0,001; p=0,023 e p=0,003). Além disso, foram observadas nas THs correlações estatisticamente significativas entre a imunoexpressão da IL-17 e IL-23 tanto nas células foliculares/tumorais, como nos linfócitos (r=0,539; p=0,038 e r=0,522; p=0,046). Sugere-se que a IL-17 e IL-23 influenciam a patogênese destas lesões tireoidianas e que em algumas delas, represente atuação da resposta Th17 (AU).

The thyroid is an endocrine gland that can be affected by several lesions, including reactive, autoimmune and neoplastic diseases. Evidence shows that the immune response, from the release of inflammatory cytokines by immune and non-immune cells, plays an important role in the development of several diseases. Among these cytokines, IL-17 and IL-23 stand out, which have been associated with the Th17 response and pathogenesis of several thyroid diseases. Hence, this research aimed to evaluate the immunoexpression of IL-17 and IL-23 proteins in papillary thyroid carcinomas (PTCs), follicular adenomas (FAs), colloid goiters (CGs) and Hashimoto's thyroiditis (HTs) (isolated and associated with PTCs or GCs), in order to better understand the role of these cytokines in these entities. The sample consisted of 30 cases of PTCs, 10 FAs, 15 CGs and 15 HTs. The analysis of protein immunoexpression in follicular/tumor cells was semiquantitatively performed in all lesions, which were classified into three scores: 1 (≤ 33%), 2 (> 33% - 66%) and 3 (> 66%). For the evaluation of immunopositive lymphocytes, five fields with the greater number of positive lymphocytes were selected, where the count was performed. The data were submitted to statistical analysis using the Kruskal-Wallis (KW), Mann-Whitney (U) and Spearman (r) tests, with the significance level set at 5% (p < 0,05). In the analysis of IL-17 between lesions, a higher immunoexpression was verified in the HT lymphocytes compared to FAs and CGs (p=0.041 and p=0.013). In the evaluation of IL-23, a higher immunostaining of follicular/tumor cells in HTs was detected in relation to PTCs (p=0.002), as well as, a greater expression of this cytokine was observed in the lymphocytes of HTs compared to PTCs, FAs and CGs (p=0.001; p=0.023 and p=0.003). In addition, statistically significant correlations between the immunoexpression of IL17 and IL-23 were observed in both follicular/tumor cells and lymphocytes (r=0.539; p=0.038 and r=0.522; p=0.046). It is therefore suggested that IL-17 and IL-23 influence the pathogenesis of these thyroid lesions and that in some of them, they represent the Th17 response (AU).

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Humanized NOD/SCID/IL-2 receptor γ-chainnull (huNSG) mice recapitulate some features of human T-cell populations that can be exploited in basic and pre-clinical research. CXCR5+ T CD8+ T-cells play an important role in the control of viral infections and tumors. Indeed, they have been associated with low-level HIV replication, making them a possible novel correlate of protection, and potentially useful in the eradication of HIV reservoirs. Here, by flow cytometry, we evaluated the reconstitution of CXCR5+ CD8+ T-cells in huNSG mice engrafted with CD34+ hematopoietic stem cells. This population was readily generated in huNSG mice, and where particularly confined to spleen and lymph nodes. These cells exhibited a follicular-like phenotype, with expression of Programmed Death (PD)-1, Inducible T-cell costimulatory (ICOS), and absence of CCR7. Moreover, CXCR5+ CD8+ T-cells had a higher expression of interleukin (IL)-21 and a higher cytotoxic potential compared with CXCR5- cells. HIV infection did not affect the frequencies of CXCR5+ CD8+ T-cells in secondary lymphoid organs. Finally, taking advantage of the high proportion of naïve T-cells in huNSG mice, we evaluated the in vitro response of splenic T-cells to the follicular profile-polarizing cytokines Transforming Growth Factor (TGF)-ß1 and IL-23. After in vitro treatment, there was an increase in CXCR5+ CD8+ T-cells, which exhibited high levels of PD-1, CD40 L and low expression of CCR7. Thus, there is a reconstitution of CXCR5+ CD8+ T-cells in huNSG mice, supporting the use of this model for exploring the biology and role of this cell population in healthy and diseased conditions.

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Dendritic cells (DC) have the unique ability to capture microorganisms and activate naive T lymphocytes. Obtaining DC derived from progenitors demands high cost and prolonged cultivation. Different immortalized DC has been isolated but most of them have immature phenotype and depending on growing factors or other stimuli to be used. In this study we characterized the cell line AP284 as a DC. AP284 cells express high levels of CD11b, MHC class II, 33D1 and CD209b. They also express high amounts of CD80 costimulatory molecule and different toll like receptors (TLR). After stimuli with TLR agonist they produce surprising amount of IL-12p40 related to IL-23 formation but not IL-12p70. They are also able to produce IL-6 and favor amplification of a Th17 but not Th1 profile. This DC line may be useful for a better understanding of factors and cellular interactions responsible for the induction of IL-12p40, IL-23 and Th17 generation.

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Paracoccidioidomycosis (PCM), a chronic granulomatous disease caused by the thermally dimorphic fungus Paracoccidioides brasiliensis and Paracoccidioides lutzii, has the highest mortality rate among systemic mycosis. The T helper 1-mediated immunity is primarily responsible for acquired resistance during P. brasiliensis infection, while susceptibility is associated with Th2 occurrence. Th17 is a population of T CD4+ cells that, among several chemokines and cytokines, produces IL-17A and requires the presence of IL-1, IL-6, and TGF-ß for differentiation in mice and IL-23 for its maintenance. Th17 has been described as an arm of the immune system that enhances host protection against several bacterial and fungal infections, as Pneumocystis carinii and Candida albicans. In this study, we aimed to evaluate the Th17 immune response and the role of Th17-associated cytokines (IL-6, IL-23, and IL-17A) during experimental PCM. First, we observed that P. brasiliensis infection [virulent yeast strain 18 of P. brasiliensis (Pb18)] increased the IL-17A production in vitro and all the evaluated Th17-associated cytokines in the lung tissue from C57BL/6 wild-type mice. In addition, the deficiency of IL-6, IL-23, or IL-17 receptor A (IL-17RA) impaired the compact granuloma formation and conferred susceptibility during infection, associated with reduced tumor necrosis factor-α, IFN-γ, and inducible nitric oxide synthase enzyme expression. Our data suggest that IL-6 production by bone marrow-derived macrophages (BMDMs) is important to promote the Th17 differentiation during Pb18 infection. In accordance, the adoptive transfer of BMDMs from C57BL/6 to infected IL-6-/- or IL-17RA-/- mice reduced the fungal burden in the lungs compared to nontransferred mice and reestablished the pulmonary granuloma formation. Taken together, these results suggest that Th17-associated cytokines are involved in the modulation of immune response and granuloma formation during experimental PCM.

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OBJECTIVES: The mRNA expression profiles of IL-23/Th17 and the Treg-associated cytokine TGF-ß in peri-implantitis are currently under research. This study characterized the IL-17, IL-23, and TGF-ß gene expression levels in healthy and diseased peri-implant tissues and correlated these data with radiographic bone loss. MATERIAL AND METHODS: Peri-implant soft tissues from 40 subjects (20 healthy controls with mean age of 59.4 ± 6.3, and 20 with peri-implantitis with mean age of 56.6 ± 5.5) were enrolled in this study, and real-time PCR (RT-PCR) was used to define the profile of IL-17, IL-23, and TGF-ß gene expression. RESULTS: Higher levels of TGF-ß mRNA were observed in biopsies taken from healthy controls, and the IL-23 mRNA levels were significantly increased in the peri-implantitis group (P < 0.0001). No differences in IL-17 mRNA levels were observed between the two groups (P > 0.05). CONCLUSIONS: Data presented in this report demonstrated a predominant Th17 response in peri-implantitis subjects based on the higher levels of IL-23 and lower levels of TGF-ß detected.

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A periodontite e a doença inflamatória intestinal (DII) resultam de uma resposta imune exagerada iniciada ou não por uma agressão bacteriana. As citocinas IL-1ß, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL23, IL-25, IL-31, IL-33, IFN-γ, sCD40L e TNF-α são relevantes na via da resposta imune Th17, que desempenham um papel importante na patogênese de ambas as doenças. Porém, existe a necessidade de uma avaliação da coexistência destas citocinas. O objetivo do estudo foi avaliar a expressão das citocinas: IL1-ß, IL-4, IL-6, IL10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IFN-γ, TNF-α e do ligante solúvel do CD40 (sCD40L) no tecido gengival (G) e compará-la com a expressão na mucosa intestinal (I) de forma pareada em pacientes com periodontite crônica e DII. Participaram do estudo 22 pacientes com DII, média de idade de 40,0±14,5 anos. Foram registrados os parâmetros clínicos e periodontais. As biópsias gengivais e intestinais foram coletadas e homogeneizadas usando um disruptor celular. Os níveis das citocinas foram analisados pelo método multiensaio multiplex e posteriormente compensados pelo peso das amostras. As comparações entre as variáveis foram analisadas pelos testes Mann-Whitney e Wilcoxon. As correlações foram realizadas pelo teste de Spearman. Níveis significativamente maiores de IL-23 (p=0,02) e IFN-γ (p=0,01) e significantemente menores de IL-31 (p=0,02) e TNF-α (p=0,01) foram encontrados no tecido gengival quando comparamos com o tecido intestinal. Correlações moderadas significantes ocorreram entre o tecido gengival e o tecido intestinal de forma positiva para a IL-6 (G) com a IL-23(I) e de forma negativa entre IL-23 (G) com IL-1ß (I), IL-10 (I), IL-17A (I) e IFN-γ (I). Concluímos que a IL-23 e IFN-γ encontram-se significantemente aumentadas no tecido gengival, quando comparados ao tecido intestinal, sugerindo um papel importante destas citocinas na manifestação da periodontite em pacientes com DII. A presença de correlações significantemente moderadas negativas entre IL-23 gengival com diversas citocinas do tecido intestinal sugere a importância da IL-23 na patogênese da periodontite em pacientes com DII.

Periodontitis and inflammatory bowel disease (IBD) result from an excessive immune response initiated or not by a bacterial challenge. The cytokines IL-1ß, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL23, IL-25, IL-31, IL-33, IFN-γ, sCD40L and TNF-α are related to Th17 response and play an important role in the pathogenesis of both diseases. Therefore, there is a need to evaluate the coexistence of these cytokines. The aim of this study was to analyze the levels of IL1-ß, IL-4, IL-6, IL10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IFN-γ, TNF-α and sCD40L in the gingival tissue (G) and compare to the levels of the paired intestinal mucosa (I) in patients having both chronic periodontitis and IBD. Twenty-two IBD patients with chronic periodontitis and IBD, mean age 40.0±14.5, were enrolled in the study. Patients were assessed using anamnesis and periodontal examination. Gingival and intestinal biopsies were collected and homogenized using a cell disruptor. Cytokine's expression was evaluated through multiplex technology and then compensated by weight of samples. Mann-Whitney, Wilcoxon and Spearman correlation tests were used to analyze the data. Significant higher levels of gingival IL-23 (p=0.02) and IFN-γ (p=0.01), and significant lower levels of IL-31(p=0.02) and TNF-α (p=0.01) were found when compared to intestinal mucosa. Significant moderate correlations between gingival and intestinal tissue were noted to IL-6 (G) with IL-23(I) in a positive way and in a negative way to IL-23 (G) with IL-1ß (I), IL-10 (I), IL-17A (I) and IFN-γ (I). We conclude that IL -23 and IFN- γ are significantly increased in the gingival tissue, when compared to the intestinal tissue, suggesting an important role of these cytokines in the manifestation of periodontitis in patients with IBD. The presence of significantly moderate correlation between IL -23 gingival with various cytokines in the intestinal tissue suggest the importance of IL -23 in interrelation with periodontitis and inflammatory bowel disease.

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Psoriasis is a chronic autoimmune inflammatory disease that affects the skin and the joints. Psoriasis is characterized by the keratinocyte proliferation, which is induced by cytokines Th1 and Th17. Patients with plaque psoriasis present a chronic inflammatory response with high levels of interleukin (IL)-12 and IL-23. Various single-nucleotide polymorphisms (SNP) have been identified in the IL12B gene, such as SNP 3' UTR 1188 A/C (SNP rs3212227), which has been associated with susceptibility to developing plaque psoriasis and with the production of IL-12 and IL-23 in individuals of different ethnic groups. In this study, we determined whether there is an association of SNP rs3212227 with the susceptibility of developing plaque psoriasis and with serum levels of IL-12 and IL-23 in Mestizo population in western Mexico. We included 112 patients with psoriasis and 112 clinical healthy individuals in the study. The frequencies of genotypes A/A, A/C, and C/C in patients with plaque psoriasis were 41, 53, and 6%, respectively, while in the control group, these were 37, 53, and 10%, respectively, without finding statistically significant differences between both groups (p>0.05). Although IL-12 and IL-23 serum levels were higher in patients than in controls, we found no significant differences. The group of patients with genotype CC presented the highest levels of IL-23 (p<0.05). These data suggest that the SNP rs3212227 phenotype is not associated with the risk of developing plaque psoriasis or with IL-12 and IL-23 levels in Mestizo population in western Mexico.

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O tráfego de leucócitos é um processo complexo, dependente da ação de inúmeras substâncias químicas, além da perfeita interação celular. Desta forma, este estudo teve como objetivo avaliar a ação dos GCe e da ANXA1 sobre o eixo SDF-1α/CXCR4 e IL-17/IL-23/G-CSF e sobre a expressão de moléculas de adesão CD18, CD49d e CD62L. Foram utilizados camundongos machos Balb/C selvagens (WT) ou ANXA1-/-. As avaliações foram realizadas em condições basais, na presença de altas concentrações de GCe e na vigência de processo inflamatório, induzidos pela administração de ACTH (5 µg/animal, i.p.) ou pela injeção de LPS (100 µg/kg, i.p.), respectivamente, ou na ausência da ação dos GCe, pela ação do RU 38486 (RU, 10 mg/kg, i.p.). A participação da ANXA1 e do receptor FPR2 foi avaliada pelo pré-tratamento com Ac2-26 (1 mg/Kg, i.p.) ou com BOC2 (10 µg/animal, i.p.) durante 4 dias, 1 vez ao dia. A quantificação total e diferencial das células foi realizada em câmara de Neubauer e em esfregaços corados por May-Grunwald ou citometria de fluxo. As quantificações de CXCR2, CXCR4, FPR2, CD18, CD49d, CD62L e maturação granulocítica (CD11b/Ly6G) em células da medula e da circulação foram realizadas por citometria de fluxo. A expressão de ANXA1 nos tecidos do estomago e do baço foi realizada por western blotting e nas células da medula óssea e sangue circulante foi realizada por imunofluorescência. As quantificações de IL-17, IL-23, G-CSF, SDF-1α e corticosterona foram realizadas por ELISA. A quimiotaxia de neutrófilos da medula óssea e sangue periférico foi ensaiada na placa de quimiotaxia com filtro de poro de 8 µm. A fagocitose de neutrófilos apoptóticos por macrófagos da medula óssea foi avaliada por ensaio in vitro. Para verificar os efeitos do ACTH na migração de neutrófilos no processo inflamatório, foi empregado o modelo de bolsa de ar (100 µg/mL; LPS); e o comportamento dos leucócitos circulantes de animais tratados com ACTH foi avaliado pela técnica de microscopia intravital. Os resultados obtidos, que estão apresentados em quatro temáticas, mostraram que: 1) neutrófilos da medula óssea e sangue periférico expressam ANXA1 no citoplasma e membrana, bem como o receptor FPR2, constitutivamente, e a expressão de ambos é regulada pelos GCe. A ANXA1, via receptor FPR2 expresso em células da medula óssea, controlam a maturação neutrofílica e o tráfego destas células da medula óssea para o sangue. A ANXA1, via interação ao FPR2, controla o clearance de neutrófilos do sangue para a medula óssea, modulando o eixo SDF-1α/CXCR4; 2) A administração do ACTH causa neutrofilia e os neutrófilos circulantes são ANXA1+, CD18+, CD49d+, CD62L+, mostrando que injeção do ACTH in vivo altera o fenótipo destas células na circulação. Estas modificações alteram o comportamento dos neutrófilos na circulação, bem como a migração para a bolsa de ar na vigência de inflamação e para os tecidos de clearance. Estes efeitos podem ser dependentes, pelo menos em parte, da inibição de migração orientada, já que quimiotaxia frente ao fMLP ou ao SDF-1α estavam reduzidas. Ainda, o clearance de neutrófilos é reduzido em animais tratados com o ACTH pela menor atividade fagocítica e secretora dos macrófagos medulares; 3) Animais tratados com RU 38486 e ANXA1-/- mobilizam granulócitos da medula óssea para o sangue circulante e, deste compartimento para o foco de inflamação com maior intensidade que o observado em animais controles. O eixo IL-17/IL-23/G-CSF parece estar envolvido na granulopoiese e na mobilização de neutrófilos para o sangue durante a inflamação, mas não é alvo de ação da ANXA1 e o GCe nesta etapa do processo inflamatório. Adicionalmente, foi observado que na vigência de peritonite, as moléculas de adesão, CD49d e CD62L estão envolvidas no processo de migração de neutrófilos da medula óssea para o sangue. Os resultados aqui obtidos permitem concluir que os GCe e a ANXA1 são relevantes para granulopoiese e tráfego dos neutrófilos da medula óssea em condições fisiológicas e na vigência de processo inflamatório. Ainda, em conjunto com os dados da literatura, os nossos resultados podem sugerir a participação da ANXA1 dos GCe na plasticidade fenotípica dos neutrófilos de acordo com os estímulos a que são submetidos, e podem auxiliar na compreensão dos novos conceitos sobre a produção, tempo de vida, localização e funções de neutrófilos

The traffic leukocytes is a complex process dependent on the action of severals chemical mediators, in addition to perfect cell interaction. Therefore, this study aimed to evaluate the effect of GCe and ANXA1 on SDF-1α/CXCR4 and IL-17/IL-23/G-CSF and on the expression of adhesion molecules CD18, CD49d and CD62L. Balb/C wild type and ANXA1-/- male mice were employed. The analysis were performed at physiological conditions, in the presence of high concentrations of GCe and during of inflammatory process induced by ACTH administration (5 µg/animal, i.p.) or LPS injection (100 µg/kg, i.p.), respectively or in the absence of GCe action, by the action of RU 38486 (RU, 10 mg/kg , i.p.). The involvement of the receptor FPR2 and ANXA1 was assessed by pre-treatment with Ac2-26 (1 mg/kg, i.p.) or BOC2 (10 µg/animal, i.p.) for 4 days, once a day. The quantification of total and differential cell was performed in a Neubauer chamber and stained smears by May-Grunwald and flow cytometry. Quantification of expression of CXCR2, CXCR4, FPR2, CD18, CD49d, CD62L and granulocytic maturation (CD11b/Ly6G) in the bone marrow and circulation were performed by flow cytometry. The expression of ANXA1 on tissues was performed by western blotting and on cells from bone marrow and blood by immunocytochemistry. Quantification of IL-17, IL-23, G-CSF, SDF-1α and corticosterone were performed by ELISA. The chemotaxis of neutrophils from the bone marrow and blood was tested in the chemotaxis chamber with filter pore of 8 microns. The phagocytosis of apoptotic neutrophils by bone marrow macrophages was assessed by in vitro assay. To investigate the effects of ACTH in the migration of neutrophils in the inflammatory process, the model employed was air pouch (100 µg/ ml, LPS), and the behavior of circulating leukocytes from animals treated with ACTH were evaluated by intravital microscopy. The results obtained, which are presented in three sections, showed that: 1) neutrophils from the bone marrow and blood expressed ANXA1 in the cytoplasm and membrane, as well as FPR2, constitutively and the expression of both is regulated by GCe. The ANXA1 via FPR2 receptor expressed in bone marrow cells, controls the neutrophilic maturation and traffic of these cells from the bone marrow into the blood. The ANXA1 via interaction to FPR2 controls the clearance of neutrophils from the blood to the bone marrow by modulating the SDF-1α/CXCR4 axis; 2) the administration of ACTH induces neutrophilia and the circulating neutrophils are ANXA1+, CD18+, CD49d+ and CD62L+, showing that the injection of ACTH in vivo alters the phenotype of these cells in the blood. These modifications alter the behavior of neutrophils in the blood, as well as the migration to the air pouch in the presence of inflammation and to the tissue clearance, and these effects may be dependent, at least in part, on inhibition of migration oriented events, as chemotaxis in response to fMLP or SDF-1α were reduced. Further, the clearance of neutrophils is reduced in animals treated with ACTH due to the lower phagocytic and secretory activity of medullary macrophages; 3) Animals treated with RU 38486 and ANXA1-/- mobilize granulocytes from bone marrow into the blood, and from this compartment to the focus of inflammation with higher intensity than that observed in the control group. The axis IL-17/IL-23/G-CSF seems to be involved in granulopoiesis and mobilization of neutrophils into the blood during inflammation, but it is not the target of action of ANXA1 and GCe at this step of inflammatory process. Additionally, it was observed that in the presence of peritonitis, the adhesion molecules, CD49d and CD62L are involved in the migration of neutrophils from the bone marrow into the blood. The results obtained allow concluding that the GCe and ANXA1 are relevant to the granulopoiesis and the traffic of neutrophils from bone marrow under physiological conditions and in the presence of inflammation. Furthermore, together with literature data, the data presented here may suggest the involvement of ANXA1 the GCe in phenotypic plasticity of neutrophils according to the stimuli that are submitted, and may support to understand the new concepts of production, half-life, location and function of neutrophils

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La psoriasis es una enfermedad inflamatoria crónica de la piel mediada por células T que afecta a individuos con predisposición genética y presenta varios subtipos clínicos. Se caracteriza por la presencia de placas eritematosas bien definidas, escamosas y de bordes irregulares, que afectan fundamentalmente las regiones de los codos, las rodillas, el cuero cabelludo y el tronco. El alelo HLA-Cw6 del sistema principal de histocompatibilidad está relacionado con la presencia y severidad de la enfermedad. Desde el punto de vista fisiopatogénico, la psoriasis es una enfermedad inmune de tipo Th1, en la que es fundamental el eje IL-23/Th17. Las células Th17 producen las citocinas proinflamatorias (IL-17A, IL-17F, IL-22 e IL-26) que activan los queratinocitos y causan hiperproliferación y mayor producción de citocinas proinflamatorias y péptidos antimicrobianos, los que a su vez reclutan y activan otras células inmunes de la piel inflamada. Se produce así una amplificación de la respuesta inflamatoria que conduce a las manifestaciones clínicas de la enfermedad. El tratamiento de la psoriasis incluye agentes antiinflamatorios tópicos, fototerapia, inmunosupresores sistémicos y agentes biológicos, entre los que se encuentran las proteínas de fusión, los inhibidores del factor de necrosis tumoral alfa y los inhibidores de las interleucinas 12 y 23(AU)

Psoriasis is a T cell-mediated chronic inflammatory disease of the skin. It affects genetically predisposed individuals and presents several subtypes. It is characterized by the presence of well-defined erythematous, scaly, irregular border plaque or lesions, affecting mainly the elbows, knees, scalp, and trunk. The HLA-Cw6 allele of major histocompatibility system is related to the presence and severity of this disease. From the physiopathogenic viewpoint, psoriasis is a Th1-type immune disease in which the axle IL-23/Th17 is fundamental. Th17 cells produce proinflammatory cytokines (IL-17A, IL-17F, IL-22 and IL-26) which activate keratinocytes and cause hyperproliferation and increased production of proinflammatory cytokines and antimicrobial peptides. The latter, in turn, recruit and activate other immune cells of swollen skin. There is thus an amplification of the inflammatory response that leads to clinical manifestations of this disease. The treatment of psoriasis includes topical antiinflammatory agents, phototherapy and systemic immunosuppressive biological agents, including those which are fusion proteins, inhibitors of alpha tumor factor necrosis, and interleukin inhibitors 12 and 23(AU)

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RESUMEN

La psoriasis es una enfermedad inflamatoria crónica de la piel mediada por células T que afecta a individuos con predisposición genética y presenta varios subtipos clínicos. Se caracteriza por la presencia de placas eritematosas bien definidas, escamosas y de bordes irregulares, que afectan fundamentalmente las regiones de los codos, las rodillas, el cuero cabelludo y el tronco. El alelo HLA-Cw6 del sistema principal de histocompatibilidad está relacionado con la presencia y severidad de la enfermedad. Desde el punto de vista fisiopatogénico, la psoriasis es una enfermedad inmune de tipo Th1, en la que es fundamental el eje IL-23/Th17. Las células Th17 producen las citocinas proinflamatorias (IL-17A, IL-17F, IL-22 e IL-26) que activan los queratinocitos y causan hiperproliferación y mayor producción de citocinas proinflamatorias y péptidos antimicrobianos, los que a su vez reclutan y activan otras células inmunes de la piel inflamada. Se produce así una amplificación de la respuesta inflamatoria que conduce a las manifestaciones clínicas de la enfermedad. El tratamiento de la psoriasis incluye agentes antiinflamatorios tópicos, fototerapia, inmunosupresores sistémicos y agentes biológicos, entre los que se encuentran las proteínas de fusión, los inhibidores del factor de necrosis tumoral alfa y los inhibidores de las interleucinas 12 y 23

Psoriasis is a T cell-mediated chronic inflammatory disease of the skin. It affects genetically predisposed individuals and presents several subtypes. It is characterized by the presence of well-defined erythematous, scaly, irregular border plaque or lesions, affecting mainly the elbows, knees, scalp, and trunk. The HLA-Cw6 allele of major histocompatibility system is related to the presence and severity of this disease. From the physiopathogenic viewpoint, psoriasis is a Th1-type immune disease in which the axle IL-23/Th17 is fundamental. Th17 cells produce proinflammatory cytokines (IL-17A, IL-17F, IL-22 and IL-26) which activate keratinocytes and cause hyperproliferation and increased production of proinflammatory cytokines and antimicrobial peptides. The latter, in turn, recruit and activate other immune cells of swollen skin. There is thus an amplification of the inflammatory response that leads to clinical manifestations of this disease. The treatment of psoriasis includes topical antiinflammatory agents, phototherapy and systemic immunosuppressive biological agents, including those which are fusion proteins, inhibitors of alpha tumor factor necrosis, and interleukin inhibitors 12 and 23

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Las espondiloartritis son un grupo heterogéneo de enfermedades asociadas principalmente al complejo mayor de histocompatibilidad alelo HLA- B 27, y factores ambientales. La evidencia de esos desórdenes reflejan un origen autoinmune mediado por el sistema inmune adaptativo, en donde la composición de las lesiones inflamatorias está representada principalmente por macrófagos activados, linfocitos B y linfocitos T. El fenotipo y naturaleza de esas células T aún no están bien establecidos. Recientes estudios han demostrado que el clásico modelo de las células T CD4 efectoras Th-1/Th-2 en estas patologías debe ser reevaluado y darle espacio a las células Th-17 dentro de las patogenias inflamatorias articulares. Estudios preliminares dirigen la investigación hacia el eje IL-23/IL-17 en espondiloartropatías como una nueva propuesta. Considerar el bloqueo de las moléculas involucradas en esta vía podría ser interesante como nuevos blancos terapéuticos.

Spondyloarthritides are a heterogeneous group of diseases which are mainly associated with HLA B 27 and environmental factors. The evidence for these disorders reflects an adaptive immune system-mediated autoimmune origin where inflammatory lesion composition is mainly represented by activated macrophages, B lymphocytes and T lymphocytes. These T-cells phenotype and nature has not been well established. Recent studies have shown that the classical CD4+ Th-1/Th-2 effector-cell model should be reevaluated and Th-17 cells should be introduced in inflammatory joint pathogenesis. Preliminary studies have directed research towards the IL-23/IL-17 axis in spondyloarthropathies as a new proposal. The intervention of the molecules involved in this pathway might be interesting as new therapeutic targets.

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