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1.
J Virol ; 80(3): 1414-26, 2006 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-16415019

RESUMEN

We previously reported that soluble, stable YU2 gp140 trimeric human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein immunogens could elicit improved breadth of neutralization against HIV-1 isolates compared to monomeric YU2 gp120 proteins. In this guinea pig immunization study, we sought to extend these data and determine if adjuvant could quantitatively or qualitatively alter the neutralizing response elicited by trimeric or monomeric immunogens. Consistent with our earlier studies, the YU2 gp140 immunogens elicited higher-titer neutralizing antibodies against homologous and heterologous isolates than those elicited by monomeric YU2 gp120. Additionally, the GlaxoSmithKline family of adjuvants AS01B, AS02A, and AS03 induced higher levels of neutralizing antibodies compared to emulsification of the same immunogens in Ribi adjuvant. Further analysis of vaccine sera indicated that homologous virus neutralization was not mediated by antibodies to the V3 loop, although V3 loop-directed neutralization could be detected for some heterologous isolates. In most gp120-inoculated animals, the homologous YU2 neutralization activity was inhibited by a peptide derived from the YU2 V1 loop, whereas the neutralizing activity elicited by YU2 gp140 trimers was much less sensitive to V1 peptide inhibition. Consistent with a less V1-focused antibody response, sera from the gp140-immunized animals more efficiently neutralized heterologous HIV-1 isolates, as determined by two distinct neutralization formats. Thus, there appear to be qualitative differences in the neutralizing antibody response elicited by YU2 gp140 compared to YU2 monomeric gp120. Further mapping analysis of more conserved regions of gp120/gp41 may be required to determine the neutralizing specificity elicited by the trimeric immunogens.


Asunto(s)
Productos del Gen env/inmunología , Anticuerpos Anti-VIH/biosíntesis , Proteína gp120 de Envoltorio del VIH/inmunología , VIH-1/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Secuencia de Aminoácidos , Animales , Mapeo Epitopo , Productos del Gen env/administración & dosificación , Productos del Gen env/química , Productos del Gen env/genética , Cobayas , Antígenos VIH/administración & dosificación , Antígenos VIH/química , Antígenos VIH/genética , Proteína gp120 de Envoltorio del VIH/administración & dosificación , Proteína gp120 de Envoltorio del VIH/química , Proteína gp120 de Envoltorio del VIH/genética , Humanos , Técnicas In Vitro , Datos de Secuencia Molecular , Pruebas de Neutralización , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/inmunología , Estructura Cuaternaria de Proteína , Productos del Gen env del Virus de la Inmunodeficiencia Humana
2.
Vet Immunol Immunopathol ; 85(3-4): 179-88, 2002 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-11943319

RESUMEN

Challenge infections of calves with Pasteurella multocida were established to characterize the local inflammatory response and determine the effect of previous exposure to live bacteria on the post-challenge immune response. Experimental infections were established by intratracheal inoculation of P. multocida in both naive calves and calves that had been previously vaccinated with two subcutaneous (s.c.) injections of live bacteria. Histological, immunohistological and cytokine expression studies were performed on bronchoalveolar lavage (BAL) samples, lung parenchymal tissues and lung lymph nodes (LN). In comparison to uninfected control animals in which no lung lesions were observed, a patchy to confluent bronchopneumonia was observed following infection of naive calves, characterized by abscess formation, haemorrhage, oedema and suppurative consolidation. Cellular analysis following infection of naive animals was characterized by an influx of neutrophils in the BAL, with macrophages and dendritic cells observed in the lesion perimeter. A significant increase in the number of CD8(+) blasts expressing MHC (major histocompatibility) II was also observed in the BAL of infected calves. Decreased expression of interleukin (IL)-1 beta and increased expression of IL-8 compared to naive unchallenged controls was apparent in lung LN. In comparison, a more limited pathology was observed in vaccinated animals post-challenge, indicating partial protection conferred by the s.c. immunization with live bacteria. Studies of vaccinated animals showed the presence of bronchial-associated lymphoid tissue (BALT) in the lung tissue and an increase in the number of B-cells and CD4(+) T-cells expressing MHCII in the lung LN after challenge. In contrast to primary infection, there was no significant influx of neutrophils in the BAL. Instead, a population of newly recruited monocytes/macrophages was observed. Increased IL-2 expression and decreased IL-8 expression was observed in the LN, while IL-1 beta expression was not detected. The reduced neutrophil and increase monocyte response in the vaccinated calves may be associated with significant changes in the gamma delta T lymphocyte population in the BAL.


Asunto(s)
Enfermedades de los Bovinos/inmunología , Inmunización/veterinaria , Enfermedades Pulmonares/veterinaria , Infecciones por Pasteurella/veterinaria , Pasteurella multocida/crecimiento & desarrollo , Animales , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Bovinos , Enfermedades de los Bovinos/metabolismo , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/patología , Citocinas/análisis , Citocinas/biosíntesis , Citocinas/genética , Citometría de Flujo/veterinaria , Genes MHC Clase I/inmunología , Enfermedades Pulmonares/inmunología , Enfermedades Pulmonares/metabolismo , Enfermedades Pulmonares/microbiología , Neutrófilos/inmunología , Infecciones por Pasteurella/inmunología , Infecciones por Pasteurella/metabolismo , Infecciones por Pasteurella/microbiología , ARN/química , ARN/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria
3.
Immunology ; 100(1): 63-9, 2000 May.
Artículo en Inglés | MEDLINE | ID: mdl-10809960

RESUMEN

Interleukin-16 (IL-16) acts as a chemoattractant for CD4+ cells, as a modulator of T-cell activation, and plays a key role in asthma. This report describes the cytokine-inducing effects of IL-16 on total peripheral blood mononuclear cells (PBMC) and PBMC subpopulations. While CD4+ T lymphocytes did not secrete cytokines in response to rhIL-16, CD14+ CD4+ monocytes and maturing macrophages secrete IL-1beta, IL-6, IL-15 and tumour necrosis factor-alpha (TNF-alpha) upon rhIL-16 stimulation. The mRNA species for these four cytokines were detected as early as 4 hr post-stimulation, with protein being secreted by 24 hr. Secretion of IL-1beta and IL-6 by total PBMC was dose dependent, with maximal secretion being observed using 50 ng/ml rhIL-16. However, for IL-15 or TNF-alpha maximal secretion by total PBMC occurred with all concentrations between 5 ng/ml to 500 ng/ml rhIL-16. Purified monocytes/macrophages secreted maximal concentrations of all four cytokines in the presence of 500 ng/ml rhIL-16, except for monocytes where maximal secretion of IL-15 was, interestingly, observed with only 50 ng/ml rhIL-16. The use of higher concentrations of rhIL-16 (1000 ng/ml) inhibited secretion of all four cytokines. While these IL-16-induced cytokines are likely to be involved in the immune system's response to antigen, the data suggest that IL-16 may play a key role in initiating and/or sustaining an inflammatory response.


Asunto(s)
Citocinas/metabolismo , Interleucina-16/inmunología , Monocitos/inmunología , Antígenos CD4/análisis , Linfocitos T CD4-Positivos/inmunología , Técnicas de Cultivo de Célula , Diferenciación Celular/inmunología , Citocinas/biosíntesis , Citocinas/genética , Humanos , Receptores de Lipopolisacáridos/análisis , Macrófagos/inmunología , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
4.
J Immunol ; 164(9): 4429-32, 2000 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-10779741

RESUMEN

IL-16 functions as a chemoattractant factor, inhibitor of HIV replication, and inducer of proinflammatory cytokine production. Previous studies have suggested that CD4 is the receptor for IL-16, because only CD4+ cells respond to IL-16 and both the anti-CD4 Ab OKT4 and soluble CD4 can block IL-16 function. However, these are only indirect evidence of a requirement for CD4, and to date a direct interaction between IL-16 and CD4 has not been shown. In this paper, we report that cells from CD4 knockout mice are as responsive to IL-16 as their CD4 wild-type equivalents in both assays testing for IL-16 function (chemotaxis and production of proinflammatory cytokines). In addition, the inhibitory effect of soluble CD4 on IL-16 function observed using CD4 wild type murine cells was not observed using CD4 knockout cells. These data demonstrate that CD4 is not required for IL-16 function and suggest that another molecule acts as the major receptor.


Asunto(s)
Antígenos CD4/fisiología , Citocinas/metabolismo , Interleucina-16/fisiología , Animales , Antígenos CD4/genética , Células Cultivadas , Quimiotaxis de Leucocito/inmunología , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Leucocitos Mononucleares/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Monocitos/inmunología , Monocitos/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo
5.
Am J Vet Res ; 58(9): 969-75, 1997 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-9285000

RESUMEN

OBJECTIVE: To characterize cytokine profiles and lymphocyte subpopulations in lung parenchyma and bronchoalveolar lavage (BAL) fluid from normal bovine lungs. ANIMALS: Eight 12- to 18-month-old cattle. PROCEDURE: Cell populations in BAL fluid and collagenase-digested lung parenchyma were analyzed by flow cytometry and monoclonal antibodies. Proportions of total cell populations were determined, using Giemsa-stained cytospots. Distribution of lymphocytes within the lung parenchyma was analyzed by immunohistochemistry, and cytokine mRNA species in the parenchyma were characterized by use of reverse transcriptase-polymerase chain reaction analysis. RESULTS: Cytokine profiles indicated high amounts of mRNA for interleukins 6 and 10 and transforming growth factor beta. In the BAL fluid and lung parenchyma, macrophages were the predominant cell type, although the proportion was lower in the parenchyma. Lymphocytes made up approximately 3% of both cell populations. Common to both lung compartments was the predominance of CD2+ and gamma delta T cells over B lymphocytes. There were more CD8+ T cells than CD4+ T cells in both compartments. The gamma delta cells made up approximately 9% of the lymphocyte populations. Two-color flow cytometry revealed CD8+ gamma delta T cell and CD8+CD5- populations that were unique to BAL fluid. In the BAL fluid and parenchyma, most CD4+ and CD8+ T cells expressed high amounts of CD44, a characteristic of memory T cells. The gamma delta T cells were CD44(10), as were B cells in the lung parenchyma. The B cells from BAL fluid expressed high amounts of CD44. Immunohistologic analysis of lung tissue revealed bronchus-associated lymphoid tissue structures with distinctive germinal center organization of B cells encompassed by CD4+ T cells. CONCLUSIONS: Results provided normal values for comparison with those of other species and with the bovine respiratory tract response to disease.


Asunto(s)
Subgrupos de Linfocitos B/inmunología , Citocinas/biosíntesis , Pulmón/inmunología , Subgrupos de Linfocitos T/inmunología , Animales , Subgrupos de Linfocitos B/citología , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Antígenos CD2/análisis , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Bovinos , Citocinas/análisis , Cartilla de ADN , Citometría de Flujo/métodos , Receptores de Hialuranos/análisis , Pulmón/citología , Reacción en Cadena de la Polimerasa , Receptores de Antígenos de Linfocitos T gamma-delta/análisis , Valores de Referencia , Subgrupos de Linfocitos T/citología
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