RESUMEN
We previously reported a chromatography system for purifying immunoglobulin M (IgM) using N,N,N',N'-ethylenediaminetetrakis(methylenephosphonic acid)-modified zirconia particles that selectively absorb immunoglobulins. Here, we report a simple procedure for preparing biotinylated IgM from hybridoma culture medium using this zirconia-based chromatography system. The culture medium of an IgM-producing hybridoma cell line was used as the starting sample solution, and the IgM in the medium was concentrated and partially purified by zirconia chromatography. Next, 9-(biotinamido)-4,7-dioxanonanoic acid N-succinimidyl ester was added to react with the proteins in the sample. Subsequently, only the biotinylated IgM was isolated by Capto Core 400 polishing column chromatography. The entire process was easy to perform, could be completed within 2 h, and provided highly pure biotin-labeled IgM. This procedure is expected to be applicable to the labeling of IgM with various compounds and drugs.
Asunto(s)
Biotinilación , Medios de Cultivo , Hibridomas , Inmunoglobulina M , Inmunoglobulina M/química , Inmunoglobulina M/aislamiento & purificación , Animales , Medios de Cultivo/química , Ratones , Circonio/química , Biotina/químicaRESUMEN
Sporotrichosis diagnosis involves a series of analyses, including culture and antibody detection in serum samples. Serologic methods may sometimes yield false-negative or false-positive results, leading to inaccurate diagnoses. This study assessed specific patient groups in which antibody detection of different isotypes and subclasses may lack sensitivity. An enzyme-linked immunosorbent assay (ELISA) with Sporothrix brasiliensis exoantigens was used to investigate IgM, IgG, IgG1, IgG2, IgG3, IgG4, IgA, IgA1 and IgA2 antibodies in human serum samples. Eighty serum samples from patients with different sporotrichosis clinical manifestations, including cutaneous forms with and without hypersensitivity manifestations, extracutaneous forms (bone, ocular, meningeal and pulmonary), disseminated cutaneous forms and disseminated forms in individuals living with HIV/AIDS, diabetics and alcoholics, were evaluated. The ELISA sensitivities in the detection of different antibodies ranged from 0.85 to 0.60 for the detection of IgG2 and IgG3, respectively. The antibodies with higher area under ROC curves were IgG2, IgG, IgA and IgA1. There were no significant differences in the immunological reactivity of the tested antibodies among different clinical forms of sporotrichosis. The data revealed a higher likelihood of a false-negative outcome in patients with lesions in the nasal mucosa regarding the detection of IgM and a lower likelihood in patients with lymphocutaneous sporotrichosis regarding the detection of IgG3. Patients with hypersensitivity manifestations had a 3.71 odds ratio to yield negative results in total IgG detection. In conclusion, we identified specific patient groups in which antibody detection may lack sensitivity, thus contributing to a better understanding of the diagnostic challenges associated with this condition.
Asunto(s)
Anticuerpos Antifúngicos , Ensayo de Inmunoadsorción Enzimática , Sensibilidad y Especificidad , Sporothrix , Esporotricosis , Humanos , Esporotricosis/inmunología , Esporotricosis/diagnóstico , Anticuerpos Antifúngicos/sangre , Sporothrix/inmunología , Sporothrix/clasificación , Masculino , Femenino , Adulto , Persona de Mediana Edad , Isotipos de Inmunoglobulinas/sangre , Isotipos de Inmunoglobulinas/inmunología , Inmunoglobulina G/sangre , Anciano , Adulto Joven , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/sangre , Inmunoglobulina A/sangre , Inmunoglobulina M/sangreRESUMEN
Limited knowledge exists on the quality of polyclonal antibody responses generated following Marburg virus (MARV) infection and its evolution in survivors. In this study, we evaluate MARV proteome-wide antibody repertoire longitudinally in convalescent phase approximately every six months for five years following MARV infection in ten human survivors. Differential kinetics were observed for IgM vs IgG vs IgA epitope diversity, antibody binding, antibody affinity maturation and Fc-receptor interaction to MARV proteins. Durability of MARV-neutralizing antibodies is low in survivors. MARV infection induces a diverse epitope repertoire with predominance against GP, VP40, VP30 and VP24 that persisted up to 5 years post-exposure. However, the IgM and IgA repertoire declines over time. Within MARV-GP, IgG recognize antigenic sites predominantly in the amino-terminus, wing domain and GP2-heptad repeat. Interestingly, MARV infection generates robust durable FcɣRI, FcɣRIIA and FcɣRIIIA IgG-Fc receptor interactions. Immunization with immunodominant MARV epitopes reveals conserved wing region between GP1 and GP2, induces neutralizing antibodies against MARV. These findings demonstrate that MARV infection generates a diverse, long-lasting, non-neutralizing, IgG antibody repertoire that perturbs disease by FcɣR activity. This information, along with discovery of neutralizing immunogen in wing domain, could aid in development of effective therapeutics and vaccines against Marburg virus.
Asunto(s)
Anticuerpos Neutralizantes , Anticuerpos Antivirales , Enfermedad del Virus de Marburg , Marburgvirus , Proteoma , Marburgvirus/inmunología , Humanos , Enfermedad del Virus de Marburg/inmunología , Enfermedad del Virus de Marburg/virología , Anticuerpos Antivirales/inmunología , Anticuerpos Neutralizantes/inmunología , Proteoma/inmunología , Femenino , Vacunas Virales/inmunología , Inmunoglobulina G/inmunología , Masculino , Epítopos/inmunología , Adulto , Inmunoglobulina M/inmunología , Persona de Mediana Edad , Estudios Longitudinales , Inmunoglobulina A/inmunología , Desarrollo de Vacunas , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
BACKGROUND: Ankylosing spondylitis (AS) has been known to have auto-inflammatory nature; hence, the efficacy of autoantibodies is low. However, studies on autoantibodies are ongoing, with some studies showing associations. Previous studies showed that anti-protein phosphatase magnesium-dependent 1A (PPM1A) IgG was increased in patients with AS and associated with radiographic progression. However, the diagnostic usefulness was limited due to relatively low sensitivity and specificity. This pilot study evaluated the diagnostic utility of anti-PPM1A-IgM and anti-PPM1A-IgG in patients with active AS. METHODS: Serum samples were obtained from the registry cohort of a single tertiary center in Korea. Serum levels of anti-PPM1A-IgG/IgM were measured by direct ELISA. Receiver operating characteristic (ROC) analysis was used to predict the diagnostic sensitivity and specificity of serum anti-PPM1A-IgG/IgM. RESULTS: Samples were collected from 28 patients with active AS, 16 healthy controls (HCs), and 28 patients with rheumatoid arthritis (RA). Although total serum IgM was lower in the RA and AS groups than in the HC group, anti-PPM1A-IgM was significantly lower in the AS group than in the other groups. In evaluating the diagnostic utility of anti-PPM1A-IgG/IgM for AS patients compared with HCs, the area under the curve (AUC) of anti-PPM1A-IgM was 0.998 (sensitivity 96.4%, specificity 100.0%). When ROC analysis of anti-PPM1A-IgM for AS patients compared with RA patients was conducted, sensitivity was 78.6% and specificity was 71.4%, with an AUC of 0.839. CONCLUSION: Decreased anti-PPM1A-IgM levels in AS patients suggests a potential role for anti-PPM1A-IgM in the diagnosis of active AS.
Asunto(s)
Autoanticuerpos , Biomarcadores , Inmunoglobulina M , Proteína Fosfatasa 2C , Curva ROC , Sensibilidad y Especificidad , Espondilitis Anquilosante , Humanos , Espondilitis Anquilosante/sangre , Espondilitis Anquilosante/diagnóstico , Masculino , Biomarcadores/sangre , Femenino , Adulto , Inmunoglobulina M/sangre , Autoanticuerpos/sangre , Proteína Fosfatasa 2C/sangre , Persona de Mediana Edad , Inmunoglobulina G/sangre , Estudios de Casos y Controles , Artritis Reumatoide/sangre , Artritis Reumatoide/diagnóstico , Proyectos Piloto , Ensayo de Inmunoadsorción EnzimáticaRESUMEN
A core component of every blood program is the supply of safe blood and blood products. The elevated risk of transmission through these products is due to parvovirus B19 (B19V) resistance to the virus inactivation procedures. Our study aimed to screen asymptomatic blood donors for B19V at a tertiary care hospital in Chennai, Tamil Nadu, between September 2020 and June 2021. Sera from 106 healthy blood donors who tested negative for Human immunodeficiency virus (HIV), Hepatitis B surface antigen (HBsAg), Hepatitis C virus (HCV), syphilis, and malaria were tested for anti-B19V IgM and IgG using a qualitative indirect enzyme-linked immunosorbent assay (ELISA). In the study population, 23.5% (n = 25) of donors tested IgM positive, 38.6% (n = 41) tested IgG positive, and 7.5% (n = 8) tested positive for both IgM and IgG. A proportion of 61.3% (n = 65) of the blood donors tested IgG negative, suggesting they had no past B19V infection. B19V DNA was not detected in any of the subjects. The high seroprevalence of IgM indicates that blood donors may have been recently exposed to B19V, potentially posing a risk to immunocompromised individuals and those with hematological stress. Further longitudinal studies with a larger sample size are recommended to better understand the risk of B19V transfusion transmission.
Asunto(s)
Anticuerpos Antivirales , Donantes de Sangre , Inmunoglobulina G , Inmunoglobulina M , Parvovirus B19 Humano , Humanos , India/epidemiología , Parvovirus B19 Humano/inmunología , Masculino , Adulto , Femenino , Anticuerpos Antivirales/sangre , Inmunoglobulina M/sangre , Inmunoglobulina G/sangre , Estudios Seroepidemiológicos , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/sangre , Infecciones por Parvoviridae/inmunología , Persona de Mediana Edad , Adulto Joven , AdolescenteRESUMEN
Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by vascular thrombosis and obstetric morbidity, with accurate laboratory examination of antiphospholipid antibodies (aPLs) being crucial for diagnosis. This study focused on anti-ß2 glycoprotein I (aß2GPI) antibodies and aimed to establish the first population-based cutoff values for aß2GPI IgA/IgM/IgG antibodies in non-pregnant women of reproductive age in Southwest China. The study cohort comprised 181 healthy women of reproductive age for study. Blood samples were collected on an early morning fast. Anti-ß2GPI antibodies including IgA, IgM and IgG were measured in serum using the HOB® BioCLIA kit. According to the Clinical and Laboratory Standards Institute (CLSI) guidelines, the study used non-parametric percentile methods to calculate the 95th, 97.5th, and 99th percentiles cutoff values for aß2GPI IgA/IgM/IgG antibodies, along with corresponding 90% confidence intervals (CI), while excluding outliers. A total of 168 independent samples were collected for verification, including 85 samples from healthy subjects and 83 samples from APS patients, in order to evaluate the analytical performance of the obtained cutoff values. The 99th percentile cutoff values were 3.36 RU/mL for aß2GPI IgA, 27.54 RU/mL for aß2GPI IgM and 1.81 RU/mL for aß2GPI IgG, which indicated that the levels of aß2GPI IgM antibodies were generally higher compared to those of IgA and IgG antibodies. Our established reference range was confirmed to be successful in validating the detected values of aß2GPI antibodies in all healthy controls. With the 99th percentile cutoff value, the sensitivity was 14.46% for aß2GPI IgA, 22.89% for aß2GPI IgG, and 9.64% for aß2GPI IgM in APS patients. This study established population-based cutoff values that are applicable to the local population for the accurate laboratory examination of aß2GPI antibodies in non-pregnant women of reproductive age. The study also recommends paying more attention to IgM positivity in women of reproductive age.
Asunto(s)
Síndrome Antifosfolípido , Inmunoglobulina G , Inmunoglobulina M , beta 2 Glicoproteína I , Humanos , Femenino , beta 2 Glicoproteína I/inmunología , Adulto , China , Síndrome Antifosfolípido/inmunología , Síndrome Antifosfolípido/sangre , Síndrome Antifosfolípido/diagnóstico , Inmunoglobulina M/sangre , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Inmunoglobulina A/sangre , Persona de Mediana Edad , Anticuerpos Antifosfolípidos/sangre , Anticuerpos Antifosfolípidos/inmunología , Adulto Joven , Valores de Referencia , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , AdolescenteRESUMEN
Immunity protective against shigella infection targets the bacterial O-specific polysaccharide (OSP) component of lipopolysaccharide. A multivalent shigella vaccine would ideally target the most common global Shigella species and serotypes such as Shigella flexneri 2a, S. flexneri 3a, S. flexneri 6, and S. sonnei. We previously reported development of shigella conjugate vaccines (SCVs) targeting S. flexneri 2a (SCV-Sf2a) and 3a (SCV-Sf3a) using a platform squaric acid chemistry conjugation approach and carrier protein rTTHc, a 52 kDa recombinant protein fragment of the heavy chain of tetanus toxoid. Here we report development of a SCV targeting S. flexneri 6 (SCV-Sf6) using the same platform approach. We demonstrated that SCV-Sf6 was recognized by serotype-specific monoclonal antibodies and convalescent sera of humans recovering from shigellosis in Bangladesh, suggesting correct immunological display of OSP. We vaccinated mice and found induction of serotype-specific OSP and LPS IgG and IgM responses, as well as rTTHc-specific IgG responses. Immune responses were increased when administered with aluminum phosphate adjuvant. Vaccination induced bactericidal antibody responses against S. flexneri 6, and vaccinated animals were protected against lethal challenge with virulent S. flexneri 6. Our results assist in the development of a multivalent vaccine protective against shigellosis.
Asunto(s)
Anticuerpos Antibacterianos , Disentería Bacilar , Inmunoglobulina G , Antígenos O , Vacunas contra la Shigella , Shigella flexneri , Vacunas Conjugadas , Shigella flexneri/inmunología , Animales , Vacunas contra la Shigella/inmunología , Vacunas contra la Shigella/administración & dosificación , Disentería Bacilar/prevención & control , Disentería Bacilar/inmunología , Ratones , Anticuerpos Antibacterianos/sangre , Anticuerpos Antibacterianos/inmunología , Vacunas Conjugadas/inmunología , Vacunas Conjugadas/administración & dosificación , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Antígenos O/inmunología , Femenino , Ratones Endogámicos BALB C , Inmunoglobulina M/inmunología , Inmunoglobulina M/sangre , Serogrupo , Lipopolisacáridos/inmunologíaRESUMEN
OBJECTIVE: Anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) autoantibodies are one of the myositis-specific antibodies which is associated with immune-mediated necrotizing myopathy (IMNM). However, the relationship between anti-HMGCR isotypes and prognosis has not yet been fully investigated. This study was conducted to gain insight into the association between anti-HMGCR isotypes and clinical, and prognosis in IMNM patients who were positive for anti-HMGCR antibodies. METHODS: Levels of anti-HMGCR isotypes (IgG, IgA and IgM) were assessed by enzyme-linked immunosorbent assay (ELISA) in 123 consecutive serum samples obtained from 71 patients who were positive for anti-HMGCR IgG at baseline. Disease activity was assessed by manual muscle testing (MMT) 8, Physician's Global Assessment (PGA) visual analog scale (VAS), and muscle VAS. RESULTS: Baseline anti-HMGCR IgG levels were correlated with PGA VAS (r = 0.24; p = 0.04), muscle VAS (r = 0.32; p < 0.01), and MMT8(r=-0.24; p = 0.04), and baseline anti-HMGCR IgM levels were positively correlated with PGA VAS (r = 0.27, p = 0.02), muscle VAS (r = 0.24, p = 0.04). Anti-HMGCR IgM positive patients had a lower age of onset [29(25,46) vs. 51(33,65), p = 0.006], and a higher proportion of neck weakness (63.5% vs. 34.6%, p = 0.031) compared with anti-HMGCR IgM negative patients. Longitudinal analysis showed that the changes in anti-HMGCR IgG levels were correlated with the changes in the PGA VAS (ß = 3.830; p < 0.0001), muscle VAS (ß = 2.893; p < 0.0001), MMT8 (ß=-19.368; p < 0.0001), and creatine kinase (CK) levels (ß = 3900.05, p < 0.0001). Anti-HMGCR IgM levels were weakly correlated with anti-HMGCR IgA levels at baseline (r = 0.33, p < 0.01), and the variations in anti-HMGCR IgA levels were correlated with the changes in anti-HMGCR IgM levels during follow-up (ß = 0.885; p < 0.0001). There were more patients with anti-HMGCR IgM who showed a refractory course than those who were with anti-HMGCR IgM negative (polycyclic course: 40% vs. 25%; chronic continuous course: 46.7% vs. 20.5%, p = 0.018). CONCLUSION: In anti-HMGCR IgG-positive IMNM patients, the levels of anti-HMGCR IgG are associated with disease activity, and anti-HMGCR IgM is associated with refractory outcome and poor prognosis.
Asunto(s)
Autoanticuerpos , Hidroximetilglutaril-CoA Reductasas , Inmunoglobulina M , Humanos , Masculino , Femenino , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Persona de Mediana Edad , Autoanticuerpos/inmunología , Autoanticuerpos/sangre , Hidroximetilglutaril-CoA Reductasas/inmunología , Adulto , Anciano , Miositis/inmunología , Miositis/sangre , Necrosis/inmunología , Ensayo de Inmunoadsorción Enzimática , Enfermedades Musculares/inmunología , Enfermedades Musculares/sangre , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunologíaRESUMEN
BACKGROUND: Blood routine testing was the most commonly used laboratory method in clinical practice. The results are often influenced by factors such as instruments, reagents, and samples, among which, the interference of cold agglutinin is a very rare element. In our article, we reported a case of red blood cell agglutination caused by Mycoplasma pneumoniae infection. METHODS: The number of blood cells were detected by blood routine analyzer with or without treatment at 37â water bath. The red blood cell agglutination was observed through blood smear staining. The cold agglutination test were performed using O-type red blood cells added into patient's plasma and refrigerated overnight at 4â. We also used luminescent immunoassay technology to detect the content of MP antibodies in patient's serum. RESULTS: The patient's results were RBC (2.69 x 1012/L), MCH (48.5 pg), MCHC (522 g/L). Through a microscope, we observed red blood cell agglutination. The concentration of MP-igM was 60.37 AU/mL. The cold agglutination test was positive. Following a 37â water bath, the patient's results changed: RBC (3.85 x 1012/L), MCH (31.2 pg), MCHC (352 g/L). The phenomenon of massive agglutination of red blood cells has also disappeared. CONCLUSIONS: The cold agglutinin produced by MP infection can alter the results of red blood cell. During the epidemic period of MP infection, it is important to pay attention to the phenomenon of abnormal elevation of MCHC in clinical practice.
Asunto(s)
Eritrocitos , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , Neumonía por Mycoplasma/sangre , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología , Mycoplasma pneumoniae/inmunología , Crioglobulinas/análisis , Crioglobulinas/metabolismo , Masculino , Pruebas de Aglutinación , Aglutinación , Femenino , Inmunoglobulina M/sangreRESUMEN
Venous thromboembolism (VTE) is a common, deadly disease with an increasing incidence despite preventive efforts. Clinical observations have associated elevated antibody concentrations or antibody-based therapies with thrombotic events. However, how antibodies contribute to thrombosis is unknown. Here, we show that reduced blood flow enabled immunoglobulin M (IgM) to bind to FcµR and the polymeric immunoglobulin receptor (pIgR), initiating endothelial activation and platelet recruitment. Subsequently, the procoagulant surface of activated platelets accommodated antigen- and FcγR-independent IgG deposition. This leads to classical complement activation, setting in motion a prothrombotic vicious circle. Key elements of this mechanism were present in humans in the setting of venous stasis as well as in the dysregulated immunothrombosis of COVID-19. This antibody-driven thrombosis can be prevented by pharmacologically targeting complement. Hence, our results uncover antibodies as previously unrecognized central regulators of thrombosis. These findings carry relevance for therapeutic application of antibodies and open innovative avenues to target thrombosis without compromising hemostasis.
Asunto(s)
Plaquetas , COVID-19 , Activación de Complemento , Inmunoglobulina M , Trombosis , Humanos , Trombosis/inmunología , Animales , Inmunoglobulina M/inmunología , Activación de Complemento/inmunología , Ratones , Plaquetas/inmunología , Plaquetas/metabolismo , COVID-19/inmunología , COVID-19/complicaciones , SARS-CoV-2/inmunología , Proteínas del Sistema Complemento/inmunología , Proteínas del Sistema Complemento/metabolismo , Activación Plaquetaria/inmunología , Inmunoglobulina G/inmunología , MasculinoRESUMEN
SARS-CoV-2 caused the pandemic situation experienced since the beginning of 2020, and many countries faced the rapid spread and severe form of the disease. Mechanisms of interaction between the virus and the host were observed during acute phase, but few data are available when related to immunity dynamics in convalescents. We conducted a longitudinal study, with 51 healthy donors and 62 COVID-19 convalescent patients, which these had a 2-month follow-up after symptoms recovery. Venous blood sample was obtained from all participants to measure blood count, subpopulations of monocytes, lymphocytes, natural killer cells and dendritic cells. Serum was used to measure cytokines, chemokines, growth factors, anti-N IgG and anti-S IgG/IgM antibodies. Statistic was performed by Kruskal-Wallis test, and linear regression with days post symptoms and antibody titers. All analysis had confidence interval of 95%. Less than 35% of convalescents were anti-S IgM+, while more than 80% were IgG+ in D30. Anti-N IgG decreased along time, with loss of seroreactivity of 13%. Eosinophil count played a distinct role on both antibodies during all study, and the convalescence was orchestrated by higher neutrophil-to-lymphocyte ratio and IL-15, but initial stages were marked by increase in myeloid DCs, B1 lymphocytes, inflammatory and patrolling monocytes, G-CSF and IL-2. Later convalescence seemed to change to cytotoxicity mediated by T lymphocytes, plasmacytoid DCs, VEGF, IL-9 and CXCL10. Anti-S IgG antibodies showed the longest perseverance and may be a better option for diagnosis. The inflammatory pattern is yet present on initial stage of convalescence, but quickly shifts to a reparative dynamic. Meanwhile eosinophils seem to play a role on anti-N levels in convalescence, although may not be the major causative agent. We must highlight the importance of immunological markers on acute clinical outcomes, but their comprehension to potentialize adaptive system must be explored to improve immunizations and further preventive policies.
Asunto(s)
Anticuerpos Antivirales , COVID-19 , Convalecencia , Citocinas , Inmunoglobulina G , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/sangre , Masculino , Femenino , Adulto , Persona de Mediana Edad , SARS-CoV-2/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Citocinas/sangre , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Estudios Longitudinales , Anciano , Eosinófilos/inmunología , Eosinófilos/metabolismoRESUMEN
Cattle produce Abs with an H chain ultralong CDR3 (40-70 aa). These Abs have been shown to have features such as broad neutralization of viruses and are investigated as human therapeutics. A common issue in sequencing the bovine BCR repertoire is the sequence length required to capture variable (V) and isotype gene information. This study aimed to assess the use of Oxford Nanopore Technologies' MinION platform to perform IgM BCR repertoire sequencing to assess variation in the percentage of ultralong CDR3s among dairy cattle. Blood was collected from nine Holstein heifers. B cells were isolated using magnetic bead-based separation, RNA was extracted, and IgM+ transcripts were amplified using PCR and sequenced using a MinION R10.4 flow cell. The distribution of CDR3 lengths was trimodal, and the percentage of ultralong CDR3s ranged among animals from 2.32 to 20.13% in DNA sequences and 1.56% to 17.02% in productive protein sequences. V segment usage varied significantly among heifers. Segment IGHV1-7, associated with ultralong CDR3s, was used in 5.8-24.2% of sequences; usage was positively correlated with ultralong CDR3 production (r = 0.99, p < 0.01). To our knowledge, this is the first study to sequence the bovine BCR repertoire using Oxford Nanopore Technologies and demonstrates the potential for cost-efficient long-read repertoire sequencing in cattle without assembly. Findings from this study support literature describing the distribution of length and percentage of ultralong CDR3s. Future studies will investigate changes in the bovine BCR repertoire associated with age, antigenic exposure, and genetics.
Asunto(s)
Linfocitos B , Regiones Determinantes de Complementariedad , Inmunoglobulina M , Receptores de Antígenos de Linfocitos B , Animales , Bovinos , Regiones Determinantes de Complementariedad/genética , Regiones Determinantes de Complementariedad/inmunología , Receptores de Antígenos de Linfocitos B/genética , Receptores de Antígenos de Linfocitos B/inmunología , Inmunoglobulina M/inmunología , Inmunoglobulina M/sangre , Linfocitos B/inmunología , Femenino , Nanoporos , Análisis de Secuencia de ADNRESUMEN
Toxoplasmosis is one of the most widespread zoonotic diseases in the world. Human infection rates range from 10% to 80% in many countries. Female cancer patients receiving chemotherapy are more susceptible to developing acute forms of toxoplasmosis, which can cause brain defects, neurological damage, and encephalitis. The aim of the present study was to investigate the effect of Toxoplasma gondii infection on the induction of interferon-gamma in breast cancer patients from Iraq. This descriptive cross-sectional study was performed on women had breast cancer in Al-Haboubi Teaching Hospital in Nasiriya City-Thi-Qar Province (Iraq) during the period from January to September 2022. Approximately three ml of blood was drawn from all participants and sera were collected. The Sera were then tested for Toxoplasma IgM, IgG, and IFN-γ (Nova Tec Immunodiagnostica GmbH, Germany) using the enzyme-linked immunosorbent assay (ELISA) kits according to the protocols of the manufacturer. Before blood collection, participants completed a printed questionnaire with some demographic information, such as age and place of residence. The total number of positive T. gondii infections from breast cancer patients in the current study was 60 (85.7%). The results of sample analysis by ELISA assay showed that 85.7% and 74.2% of patients were positive for IgG and IgM, respectively. The mean IFN-γ levels in breast cancer patients with toxoplasmosis, without toxoplasmosis, and in the control group were 47.66, 0.00, and 0.57 pg/ ml, respectively. Higher IgG and interferon gamma levels were detected in the group of breast cancer patients with toxoplasmosis than in the group without toxoplasmosis. According to the ELISA findings, T. gondii was the most common parasite in female cancer patients.
Asunto(s)
Neoplasias de la Mama , Interferón gamma , Toxoplasma , Toxoplasmosis , Adulto , Anciano , Femenino , Humanos , Persona de Mediana Edad , Anticuerpos Antiprotozoarios/sangre , Estudios Transversales , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Interferón gamma/sangre , Interferón gamma/análisis , Irak/epidemiología , Toxoplasma/fisiologíaRESUMEN
The IgG response against SARS-CoV-2 infection can persist for over six months (long response; LR). However, among 30% of those infected, the duration can be as short as three months or less (short response; SR). The present study assembled serological data on the anti-SARS-CoV-2 IgG response duration of two previous studies and integrated these results with the plasmatic cytokine levels and genetic profile of 10 immune-relevant SNPs that were also previously published, along with the plasmatic total IgG, IgA, and IgM levels, allowing for the genetic, clinical, immunological, and epidemiological aspects of the post-COVID-19 IgG response duration to be understood. The SR was associated with previous mild acute COVID-19 and with an SNP (rs2228145) in IL6R related to low gene expression. Additionally, among the SR subgroup, no statistically significant Spearman correlations were observed between the plasma levels of IL-17A and the Th17 regulatory cytokines IFN-γ (rs = 0.2399; p = 0.1043), IL-4 (rs = 0.0273; p = 0.8554), and IL-2 (rs = 0.2204; p = 0.1365), while among the LR subgroup, weaker but statistically significant Spearman correlations were observed between the plasma levels of IL-17A and IFN-γ (rs = 0.3873; p = 0.0016), IL-4 (rs = 0.2671; p = 0.0328), and IL-2 (rs = 0.3959; p = 0.0012). These results suggest that the Th17 response mediated by the IL-6 pathway has a role in the prolonged IgG response to SARS-CoV-2 infection.
Asunto(s)
Anticuerpos Antivirales , COVID-19 , Inmunoglobulina G , Polimorfismo de Nucleótido Simple , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/epidemiología , COVID-19/sangre , COVID-19/virología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , SARS-CoV-2/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Masculino , Femenino , Receptores de Interleucina-6/genética , Persona de Mediana Edad , Adulto , Interleucina-17/sangre , Interleucina-17/genética , Citocinas/sangre , Inmunoglobulina A/sangre , Interferón gamma/sangre , Interferón gamma/genética , Inmunoglobulina M/sangre , Interleucina-4/sangre , Interleucina-4/genética , AncianoRESUMEN
BACKGROUND: Since March 2020, COVID-19 has evolved from a localized outbreak to a global pandemic. We assessed the seroprevalence of COVID-19 in three towns in the Centre Sud region of Burkina Faso. METHODS: A population-based cross-sectional survey was conducted in three middle-sized cities in Burkina Faso's Centre Sud region, from June to July 2021. Subjects aged 16 or over at the time of the survey were considered for this seroprevalence study. The Biosynex COVID-19 BSS rapid test was used to detect immunoglobulin G (IgG) and immunoglobulin M (IgM) against SARS-CoV-2. A standardized questionnaire was also administered to collect additional information. RESULTS: A total of 2449 eligible participants (age ≥ 16 years) were identified. Serological tests for COVID-19 were performed in 2155 individuals, of which 2143 valid tests were retained and analyzed. Out of the entire sample, 246 positive tests were observed, corresponding to a prevalence of 11.48%. Prevalence was 9.35% (58 cases) in Kombissiri, 12.86% (80 cases) in Manga and 11.99% (108 cases) in Pô. By gender, 13.37% of women (164 cases) tested positive, and 8.95% of men (82 cases). Women accounted for 66.67% of all positive test subjects. The results from the multivariate analysis show a significantly higher seroprevalence in women (p = 0.007), people over 55 years old (p = 0.004), overweight people (p = 0.026) and those with drinking water sources at home (p = 0.013). CONCLUSIONS: The results of this study show that the COVID-19 virus also circulates in the population of middle-sized cities in Burkina Faso, far more than officially reported by the information service of the government of Burkina Faso, given the lack of systematic testing in the general population in the country. The study also highlighted the greater vulnerability of women, older and overweight individuals to the epidemic. The preventive measures put in place to fight the pandemic must take these different factors into account.
Asunto(s)
COVID-19 , Ciudades , SARS-CoV-2 , Humanos , COVID-19/epidemiología , COVID-19/sangre , Burkina Faso/epidemiología , Femenino , Masculino , Adulto , Estudios Seroepidemiológicos , Estudios Transversales , Persona de Mediana Edad , Factores de Riesgo , Adolescente , SARS-CoV-2/inmunología , SARS-CoV-2/aislamiento & purificación , Ciudades/epidemiología , Adulto Joven , Inmunoglobulina G/sangre , Anciano , Anticuerpos Antivirales/sangre , Inmunoglobulina M/sangreRESUMEN
Hepatitis Delta Virus (HDV), a satellite virus of Hepatitis B virus, exacerbates liver damage in affected individuals. Screening for HDV antibodies in HBsAg positive patients is recommended, but the diagnostic accuracy of serological tests remains uncertain. This review aimed to assess the diagnostic accuracy of serological tests for HDV. We searched PubMed, Web of Science, Cochrane Central Register of Controlled Trials, Scopus etc. for relevant studies. Studies measuring the sensitivity and specificity of serological HDV tests against PCR as a reference standard were included. Pooled sensitivity and specificity for each test method and sero-marker were calculated. The review included six studies with 11 study arms, evaluating ARCHITECT immunoassay, EIA, ELISA, QMAC, RIA, and Western Blot test methods targeting Anti-HDV IgG, Total anti-HDV and Anti-HDV IgM. Sensitivities for Anti-HDV IgG, Total Anti-HDV and Anti-HDV IgM, tests were 97.4%, 51.9%, and 62.0%, respectively, with specificities of 95.3%, 80.0%, and 85.0%. Our findings, with its limited number of studies, suggest that HDV serological tests, particularly those identifying Anti IgG exhibit high accuracy and can serve as effective screening tools for HDV.
Asunto(s)
Hepatitis D , Virus de la Hepatitis Delta , Sensibilidad y Especificidad , Pruebas Serológicas , Humanos , Virus de la Hepatitis Delta/inmunología , Hepatitis D/diagnóstico , Hepatitis D/virología , Hepatitis D/sangre , Hepatitis D/inmunología , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Inmunoglobulina G/sangre , Anticuerpos Antihepatitis/sangre , Inmunoglobulina M/sangreRESUMEN
BACKGROUND: Toxoplasma gondii (T. gondii) is a ubiquitous protozoan parasite on our planet that causes toxoplasmosis. This study evaluated the seroprevalence and related risk factors for T. gondii infection in a population referred to healthcare centers in Meshkin-Shahr, Northwest Iran. METHODS: A total of 400 blood samples were randomly collected from the general population and assessed using the anti-Toxoplasma antibodies, Immunoglobulin G and M (IgG and IgM) Enzyme-linked immunosorbent assay (ELISA) Kits in two steps before and during the coronavirus disease 2019 (COVID-19) pandemic, 2019-2020. The results were analyzed through logistic regression via SPSS 26 software. RESULTS: Before the COVID-19 pandemic, anti-toxoplasma antibodies were detected in 39% of individuals (IgG: 38%, IgM: 0.5%, and IgG-IgM: 0.5%). Among the eleven risk factors evaluated, contact with soil and people awareness were significantly associated with T. gondii infection (p < 0.05). However, factors such as females, 20-39 age groups, junior high schools, housewives, rural areas, raw meat or vegetable consumption, vegetable or fruits washed by water, not detergent, and cat owners did not show a significant relationship with seropositivity (p > 0.05). After the outbreak of the COVID-19 pandemic, the overall seroprevalence for anti-T. gondii antibody increased to 49.7% (IgG: 47.7%, IgM: 0.5%, and IgG and IgM: 1.5%). Among these patients, 26% were positive for COVID-19. Additionally, before the COVID-19 pandemic, 40 samples were negative for anti-T. gondii antibodies but later became positive. The crude and adjusted models suggested that toxoplasmosis may be a possible risk factor for increased susceptibility to COVID-19, with an odds ratio (OR) of 1.28 (95% confidence interval (CI), 0.82-1.99; P < 0.05). Conversely, a non-significant protective effect against latent toxoplasmosis was observed in COVID-19-positive individuals (OR = 0.99; 95% CI, 0.51-1.92; P > 0.05), and COVID-19 positivity did not increase the levels of anti-T. gondii IgG antibodies. CONCLUSIONS: The general population in this region had a moderate seroprevalence of T. gondii. The increased number of COVID-19-positive patients with latent toxoplasmosis highlights the need to pay attention to the early diagnosis and proper treatment of toxoplasmosis in these patients and implement preventive programs in these areas for future possible viral infections.
Asunto(s)
Anticuerpos Antiprotozoarios , COVID-19 , Inmunoglobulina G , Inmunoglobulina M , SARS-CoV-2 , Toxoplasma , Toxoplasmosis , Humanos , COVID-19/epidemiología , COVID-19/inmunología , Irán/epidemiología , Estudios Seroepidemiológicos , Toxoplasmosis/epidemiología , Femenino , Masculino , Adulto , Toxoplasma/inmunología , Persona de Mediana Edad , Adulto Joven , Inmunoglobulina G/sangre , Factores de Riesgo , Inmunoglobulina M/sangre , Anticuerpos Antiprotozoarios/sangre , SARS-CoV-2/inmunología , Adolescente , Anciano , Niño , Prevalencia , Ensayo de Inmunoadsorción Enzimática , Preescolar , PandemiasRESUMEN
Point-of-care serological and direct antigen testing offers actionable insights for diagnosing challenging illnesses, empowering distributed health systems. Here, we report a POC-compatible serologic test for Lyme disease (LD), leveraging synthetic peptides specific to LD antibodies and a paper-based platform for rapid, and cost-effective diagnosis. Antigenic epitopes conserved across Borrelia burgdorferi genospecies, targeted by IgG and IgM antibodies, are selected to develop a multiplexed panel for detection of LD antibodies from patient sera. Multiple peptide epitopes, when combined synergistically with a machine learning-based diagnostic model achieve high sensitivity without sacrificing specificity. Blinded validation with 15 LD-positive and 15 negative samples shows 95.5% sensitivity and 100% specificity. Blind testing with the CDC's LD repository samples confirms the test accuracy, matching lab-based two-tier results, correctly differentiating between LD and look-alike diseases. This LD diagnostic test could potentially replace the cumbersome two-tier testing, improving diagnosis and enabling earlier treatment while facilitating immune monitoring and surveillance.
Asunto(s)
Anticuerpos Antibacterianos , Borrelia burgdorferi , Inmunoglobulina G , Inmunoglobulina M , Enfermedad de Lyme , Sensibilidad y Especificidad , Pruebas Serológicas , Enfermedad de Lyme/diagnóstico , Enfermedad de Lyme/inmunología , Enfermedad de Lyme/sangre , Enfermedad de Lyme/microbiología , Humanos , Pruebas Serológicas/métodos , Borrelia burgdorferi/inmunología , Anticuerpos Antibacterianos/sangre , Anticuerpos Antibacterianos/inmunología , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Antígenos Bacterianos/inmunología , Aprendizaje Automático , Epítopos/inmunología , Pruebas en el Punto de Atención , Sistemas de Atención de PuntoRESUMEN
Hantavirus cardiopulmonary syndrome is a severe illness transmitted by rodent excretions. We describe a case of a 24-year-old man who presented to the emergency department with cough, shortness of breath, chills, myalgias, nausea, and diarrhea. Physical examination and laboratory analysis revealed signs of respiratory distress and thrombocytopenia. The trajectory of his illness led to acute respiratory distress syndrome (ARDS) and hemodynamic instability. Serum testing was positive for hantavirus IgM and IgG antibodies. The patient was managed with supportive care and improved. This case highlights the importance of considering hantavirus when managing patients who develop thrombocytopenia, ARDS, and hemodynamic instability in the appropriate clinical setting.
Asunto(s)
Síndrome Pulmonar por Hantavirus , Orthohantavirus , Humanos , Masculino , Síndrome Pulmonar por Hantavirus/diagnóstico , Adulto Joven , Animales , Orthohantavirus/inmunología , Trombocitopenia/etiología , Síndrome de Dificultad Respiratoria/etiología , Anticuerpos Antivirales/sangre , Inmunoglobulina G/sangre , Ratones , Inmunoglobulina M/sangreRESUMEN
PURPOSE: The current diagnostic methods for leptospirosis diagnosis are technically complex and expensive, with limited applicability to specialized laboratories. Furthermore, they lack diagnostic accuracy in the acute stage of the disease, which coincides with a period when antibiotics are highly effective. New simple and accurate tests are mandatory to decentralize and improve diagnosis. Here, we introduced a new lateral flow immunoassay (Lepto-LF) for human leptospirosis. METHODS: We conducted a double-blinded assay using 104 serum samples from patients with confirmed or discarded diagnosis for leptospirosis. The diagnostic performance of Lepto-LF was estimated across different ranges of days from onset of symptoms (dpo), considering the diagnostic algorithm as reference standard. Additionally, it was compared with the screening methods enzyme-linked immunosorbent assay (IgM-ELISA) and the slide agglutination test using temperature-resistant antigen (SATR). RESULTS: Lepto-LF exhibited perfect diagnostic performance with a Youden´s index J = 1 from 6 dpo in the acute phase. IgM-ELISA gave slightly lower accuracy with J = 0.91 and 95.5% of both sensitivity and specificity; while SATR showed a markedly inferior yield (J = 0.41, sensitivity = 95.5%, specificity = 45.5%). The performances remained consistent in the convalescence phase of the disease (> 10 dpo). CONCLUSION: Lepto-LF was found to be a reliable test for simple, rapid and early diagnosis of leptospirosis, resulting a promising tool for decentralizing leptospirosis diagnosis and enabling timely treatment of patients. In addition, Lepto-LF may be employed as confirmatory test, especially in remote areas and vulnerable contexts where the standard MAT is not available.