RESUMEN
Chronic inappropriate immune activation is the central defect-driving loss of CD4(+) T helper cells and progression to AIDS in persons with HIV-1 infection, but the mechanisms remain controversial. We examined key regulatory invariant receptor natural killer T (iNKT) cells in the gut, the largest reservoir of lymphocytes and a key arena of HIV-1 pathogenesis. In healthy control persons, the anti-inflammatory CD4(+) iNKT-cell subset predominated over the pro-inflammatory CD4(-) iNKT-cell subset in the gut, but not in the blood, compartment. HIV-1 infection resulted in a preferential loss of this anti-inflammatory CD4(+) iNKT-cell subset within the gut. The degree of loss of the CD4(+) iNKT-cell subset in the gut, but not in the blood, correlated to the systemic immune activation and exhaustion that have been linked to disease progression. These results suggest a potentially important contribution of gut iNKT-cell imbalance in determining the systemic immune activation that is the hallmark of HIV-1 pathogenesis.
Asunto(s)
Infecciones por VIH/inmunología , VIH-1/inmunología , Intestinos/inmunología , Depleción Linfocítica , Células T Asesinas Naturales/inmunología , Adulto , Antígenos CD4/metabolismo , Muerte Celular , Progresión de la Enfermedad , Humanos , Inmunomodulación , Intestinos/virología , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Células T Asesinas Naturales/virología , Activación Viral/inmunología , Adulto JovenRESUMEN
We have made thioglycoside donors for the 4,6-dideoxy-L-lyxo-hexopyranosyl ('4-deoxy-L-rhamnosyl') and 4-deoxy-4-fluoro-L-rhamnosyl monosaccharide residues. The preparation of the deoxyfluororhamnose was not straightforward, and revealed some unexpected behavior of the diethylaminosulfur trifluoride (DAST) reagent. The new glycosyl donors were used to synthesize two analogs of the mycobacterial arabinogalactan linkage disaccharide -->4)-alpha-L-Rha-(1-->3)-alpha-D-GlcNAc. These analogs are prototypes for a family of potential inhibitors of the enzymes involved in the early stages of cell-wall construction in mycobacteria.
Asunto(s)
Disacáridos/síntesis química , Galactanos/química , Glucósidos/síntesis química , Mycobacterium/química , Conformación de Carbohidratos , Dietilaminas/química , Flúor/química , Compuestos de Flúor/síntesis química , Espectroscopía de Resonancia Magnética , Ramnosa/análogos & derivados , Tioglicósidos/síntesis químicaRESUMEN
OBJECTIVE: To evaluate the efficacy of combination protease and reverse transcriptase inhibitor therapy in correcting HIV-1-induced lymphocyte subset abnormalities in previously treated adults. DESIGN: A 48-week observational study of lymphocyte subsets in 12 participants in the Multicenter AIDS Cohort Study who were already taking at least one reverse transcriptase inhibitor and added a protease inhibitor to their treatment regimen. Comparison groups were HIV-seronegative homosexual men, HIV-seronegative heterosexual men, and homosexual HIV-1-infected men who were long-term non-progressors. METHODS: Three-color immunofluorescence and monoclonal antibodies were used to assess HIV-1-induced lymphocyte subset alterations related to immune deficiency and immune activation. Plasma HIV-1 RNA levels were monitored to assess suppression of viral replication. RESULTS: CD4+ cell counts significantly increased and lymphocyte activation measured as CD38 and HLA-DR expression on CD8+ T cells significantly decreased by 48 weeks. CD4+ cell values remained abnormal even in those who were fully suppressed. Some T-cell activation markers decreased to levels observed in long-term non-progressors. The increase in CD4+ T-cell numbers reached a plateau by week 24, but the increase in resting HLA-DR- CD38-T cells was sustained through week 48. Proportions of CD45RA+ CD62L-selectin+ and CD28+ CD4+ T-cell subsets and Fas expression were not abnormal at baseline compared with seronegative homosexual controls. CONCLUSIONS: The most significant impact of suppression of viral replication was reversal of T-cell activation. However, normalization of lymphocyte subset perturbations associated with chronic HIV-1 infection was not achieved after 1 year of treatment with current combination antiretroviral regimens. More profound viral suppression, therapy for longer than 1 year, or immunologic augmentation may be needed to fully reverse the abnormalities.
Asunto(s)
Fármacos Anti-VIH/uso terapéutico , Infecciones por VIH/inmunología , Inhibidores de la Proteasa del VIH/uso terapéutico , VIH-1 , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Adulto , Anticuerpos Monoclonales , Recuento de Linfocito CD4 , Enfermedad Crónica , Estudios de Cohortes , Quimioterapia Combinada , Técnica del Anticuerpo Fluorescente , Infecciones por VIH/tratamiento farmacológico , Sobrevivientes de VIH a Largo Plazo , Seronegatividad para VIH , Homosexualidad Masculina , Humanos , Activación de Linfocitos , Masculino , ARN Viral/sangre , Subgrupos de Linfocitos T/inmunología , Replicación ViralRESUMEN
A trimeric beta-lactosyl cluster based on 2-nitro-2-(hydroxymethyl)propane-1,3-diol was prepared using the trichloroacetimidate method. Kinetic studies showed that this cluster was an effective acceptor for rat-liver alpha-(2-->3)-sialyltransferase. Its KM was comparable to those for monomeric lactose and N-acetyllactosamine acceptors, and its Vmax was 1% of that measured for the LacNAc acceptor. Preparative-scale sialylation using this enzyme afforded a trimeric cluster of the ganglioside GM3 oligosaccharide in good yield. The NMR spectra of the trimeric GM3 analogue suggest that the oligosaccharide conformation is not significantly perturbed by this level of clustering.
Asunto(s)
Gangliósido G(M3)/análogos & derivados , Gangliósido G(M3)/síntesis química , Amino Azúcares/metabolismo , Animales , Conformación de Carbohidratos , Secuencia de Carbohidratos , Gangliósido G(M3)/metabolismo , Cinética , Lactosa/análogos & derivados , Lactosa/metabolismo , Hígado/enzimología , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Monosacáridos/análisis , Monosacáridos/química , Ratas , Sialiltransferasas/metabolismoRESUMEN
In experimental murine inflammation-associated amyloidosis (AA amyloidosis), an interaction between heparan sulfate and serum amyloid A (SAA), the AA precursor, has been demonstrated and is believed to play an important role in AA amyloidogenesis. Poly(vinylsulfonate) sodium salt (PVS) can arrest AA amyloid induction and cause established amyloid deposits to regress. PVS is thought to have this property by virtue of limited anionic structural similarities it has to heparan sulfate. In the present study, a comparison has been made of the in situ light microscopic and high-resolution ultrastructure of amyloid deposits before and after PVS treatment. As shown recently in situ, AA fibrils from untreated mice are composed of an outer layer of heparan sulfate proteoglycan and a 1- to 2-nm filament network of AA protein. This layer encloses a microfibril-like structure composed of chondroitin sulfate proteoglycan wound around a core of amyloid P component. After treatment with PVS, both the heparan sulfate proteoglycan and the AA filament network are lost from the fibrils, and the more central portion disintegrates into the chondroitin sulfate proteoglycan with associated amyloid P subunits. These findings add further support to the concept that heparan sulfate proteoglycan is important in amyloid fibril structure, and interference with its binding interactions with the amyloid filament protein provides a point of therapeutic attack.
Asunto(s)
Polivinilos/farmacología , Proteína Amiloide A Sérica/efectos de los fármacos , Proteína Amiloide A Sérica/ultraestructura , Ácidos Sulfónicos/farmacología , Amiloidosis/metabolismo , Amiloidosis/patología , Amiloidosis/prevención & control , Animales , Femenino , Ratones , Ratones Endogámicos , Polivinilos/uso terapéutico , Proteína Amiloide A Sérica/antagonistas & inhibidores , Bazo/química , Bazo/efectos de los fármacos , Ácidos Sulfónicos/uso terapéuticoRESUMEN
Relative fluorescence intensity measurements from a flow cytometer were used to evaluate expression of CD38 and HLA-DR antigens. These molecules are associated with cellular activation and are present at increased levels on the CD8+ lymphocytes of HIV-infected subjects. In the current study, the prognostic value of mean fluorescence intensity measurements of CD38 and HLA-DR on CD8+ cells was compared to results from our previous study in which we reported prognostic value for an elevated percentage of CD8+ cells that were positive for expression of the CD38 antigen (Giorgi et al.: JAIDS 6:904-912, 1993). Using the proportional hazards model, elevated mean fluorescence intensity of CD38 expression on CD8+ cells had prognostic value for development of AIDS that was almost identical to the prognostic value of the percentage of CD8+ cells that were positive for expression of CD38. This prognostic value was in addition to that provided by the patient's CD4+ cell measurement. To our knowledge, this is the first report that a measurement of fluorescence intensity can be used as a prognostic marker in an immunodeficiency disease. Efforts are needed to establish methods that will allow widespread application of this observation in the clinical management of HIV-infected subjects.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/sangre , Antígenos CD/análisis , Antígenos de Diferenciación/análisis , Linfocitos T CD8-positivos/química , N-Glicosil Hidrolasas/análisis , ADP-Ribosil Ciclasa , ADP-Ribosil Ciclasa 1 , Síndrome de Inmunodeficiencia Adquirida/fisiopatología , Estudios de Cohortes , Progresión de la Enfermedad , Técnica del Anticuerpo Fluorescente , Estudios de Seguimiento , Antígenos HLA-DR/análisis , Prueba de Histocompatibilidad , Humanos , Glicoproteínas de MembranaRESUMEN
Natural killer (NK) cells were enumerated by three-color immunofluorescence in 255 uninfected and 399 human immunodeficiency virus-infected adults. Several dramatic alterations were observed. First, the median number and percentage of CD16+CD56+ NK cells, the subset that comprises > 90% of the NK cells in healthy adults, were severely decreased (median, 175/mm3 in uninfected controls; 63/mm3 in HIV-infected non-AIDS subjects). Even subjects with > 800 CD4+ cells/mm3 had decreased CD16+CD56+ NK cell levels (97/mm3). Second, the number of CD16+CD56- cells, an NK population that is rare in healthy adults, was elevated (median, 20/mm3 in uninfected controls; 64/mm3 in HIV-seropositive non-AIDS subjects). Third, the expression of CD16 on the NK cells was markedly reduced; some CD56+ cells and virtually all CD56- cells were CD16dim. Fourth, fluorescence-activated cell-sorting studies revealed little NK- or antibody-dependent cellular cytotoxic activity in the CD16dimCD56- cell population. These results indicate that the pathogenesis of HIV disease includes numerical alterations in subpopulations of NK cells. A better understanding of how HIV infection causes this aspect of pathogenesis is needed.
Asunto(s)
Antígeno CD56/inmunología , Infecciones por VIH/inmunología , Células Asesinas Naturales/inmunología , Subgrupos Linfocitarios/inmunología , Receptores de IgG/inmunología , Síndrome de Inmunodeficiencia Adquirida/inmunología , Adulto , Citotoxicidad Celular Dependiente de Anticuerpos/inmunología , Antivirales/uso terapéutico , Separación Celular , Estudios de Cohortes , Citotoxicidad Inmunológica , Femenino , Citometría de Flujo , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/etiología , Seropositividad para VIH/inmunología , Humanos , Inmunofenotipificación , Recuento de Linfocitos , Masculino , Análisis de RegresiónRESUMEN
Amyloid is a term for extracellular protein fibril deposits that have characteristic tinctorial and structural properties. Heparan sulphate, or the heparan sulphate proteoglycan perlecan, has been identified in all amyloids and implicated in the earliest stages of inflammation-associated (AA) amyloid induction. Heparan sulphate interacts with the AA amyloid precursor and the beta-peptide of Alzheimer's amyloid, imparting characteristic secondary and tertiary amyloid structural features. These observations suggest that molecules that interfere with this interaction may prevent or arrest amyloidogenesis. We synthesized low-molecular-weight (135-1,000) anionic sulphonate or sulphate compounds. When administered orally, these compounds substantially reduced murine splenic AA amyloid progression. They also interfered with heparan sulphate-stimulated beta-peptide fibril aggregation in vitro.
Asunto(s)
Alcanosulfonatos/uso terapéutico , Amiloidosis/tratamiento farmacológico , Proteína Amiloide A Sérica/efectos de los fármacos , Sulfatos/uso terapéutico , Enfermedad Aguda , Alcanosulfonatos/síntesis química , Alcanosulfonatos/toxicidad , Enfermedad de Alzheimer/tratamiento farmacológico , Amiloidosis/inducido químicamente , Animales , Aniones , Enfermedad Crónica , Glicoles/síntesis química , Glicoles/uso terapéutico , Glicoles/toxicidad , Heparitina Sulfato/farmacología , Ratones , Polivinilos/química , Polivinilos/uso terapéutico , Polivinilos/toxicidad , Proteína Amiloide A Sérica/análisis , Proteína Amiloide A Sérica/metabolismo , Proteína Amiloide A Sérica/ultraestructura , Bazo/patología , Sulfatos/síntesis química , Sulfatos/toxicidadRESUMEN
We examined the T-lymphocyte phenotypes of 67 human immunodeficiency virus (HIV)-infected children (P-1 or P-2) and 65 age-matched, healthy, control children stratified into four groups from < 1 to > or = 5 years of age to determine expression of antigens associated with cell activation/differentiation. Immunophenotyping was performed by laser flow cytometry using two-color immunofluorescent labeling. Although the control children showed a decline in total CD4 cell percent with age, the HIV-infected children in all age groups showed significantly decreased CD4 cell numbers compared with the age-matched controls. However, the slope of the CD4 cell decline with age was not significantly different in HIV-infected and control children. The CD4 cell decrease in infected children was reflected in both the CD45RA+ (naive) and CD45RA- (memory) CD4 cell subsets, although the CD45RA+ cells were decreased in greater proportion. Results assessing CD4 cells for expression of the L-selectin (Leu8) molecule were similar to those for CD45RA. The overall CD8 cell percentage was significantly increased in HIV-infected children compared with controls in all age groups. This was due primarily to increases in CD8 cells that were CD38+, CD57+, HLA-DR+, or CD45RA-. In a retrospective analysis of data from 23 P-0 children, we compared phenotype results from 5 children who were HIV+ with those 18 who were HIV-. Although the phenotypic changes seen in the 5 HIV+ children paralleled those described above for P-1 and P-2 subjects, there was no significant difference in the values for HIV+ compared with HIV P-0 children. Although the phenotypic alterations described did not appear to be diagnostic markers in P-0 children, they may serve as useful adjuncts for the evaluation of HIV-infected children.
Asunto(s)
Antígenos CD/sangre , Infecciones por VIH/inmunología , Subgrupos de Linfocitos T/inmunología , ADP-Ribosil Ciclasa , ADP-Ribosil Ciclasa 1 , Envejecimiento/inmunología , Antígenos de Diferenciación/sangre , Antígenos de Diferenciación de Linfocitos T/sangre , Antígenos CD4/sangre , Linfocitos T CD4-Positivos/inmunología , Antígenos CD57 , Antígenos CD8/sangre , Niño , Preescolar , Citometría de Flujo , Antígenos HLA-DR/sangre , Humanos , Inmunofenotipificación , Lactante , Recién Nacido , Antígenos Comunes de Leucocito/sangre , Recuento de Leucocitos , Glicoproteínas de Membrana , Análisis de Regresión , Estudios Retrospectivos , Linfocitos T Reguladores/inmunologíaRESUMEN
Despite the previous description of the leukocyte differentiation antigen CD20 as B cell restricted, the findings reported here indicate that a small subset of human T cells expresses low levels of CD20 or a cross-reacting antigen. Three different CD20 monoclonal antibodies (mAb), Leu16, B1, and 1F5, reacted with the T cell subset. B cells that expressed CD20 were CD20bright and constituted an average of 9.2 +/- 3.3% of adult PBL. Meanwhile, T cells that expressed CD20 were CD20dim and represented 2.4 +/- 1.5% of the PBL. This population may have been overlooked in previous studies due to the low level of CD20 expression per T cell and the small size of the subset in most individuals. Blocking studies indicated that CD20 mAb binding to CD3+ cells was due to the antigen-reactive regions of the CD20 antibodies and was not a result of Fc receptor binding, or non-specific fluorochrome or protein binding. The T cell nature of the CD20dim CD3+ cells was confirmed by the rapid rise in the intracellular calcium concentration ([Ca2+]i) of CD20dim cells observed following treatment with CD3 mAb but not following treatment with anti-human immunoglobulin (Ig). Extensive three-color immunophenotypic analyses indicated that CD20dim T cells were phenotypically heterogeneous and displayed a leukocyte differentiation profile that was slightly different than that of CD20- T cells. Thus, the CD20dim T cells were more likely than CD20- T cells to be gamma/delta T cell antigen receptor positive (14% vs. 3.4%), CD8+ (57% vs. 33%), and CD45RO+ (82% vs. 51%); fewer were CD38+ (5% vs. 24%) or CD4+ (35% vs. 61%).(ABSTRACT TRUNCATED AT 250 WORDS)