RESUMEN
Interferon-lambda receptor 1 (IFNLR1) is the key to interferon-lambda's biological activities. Rhesus macaques (Macaca mulatta) are supposedly more suitable for translational studies on interferon lambda-associated human diseases, yet little is known about their IFNLR1 (mmuIFNLR1). In this study, we cloned the coding sequence of mmuIFNLR1, examined its variants, and determined the distribution of mmuIFNLR1 mRNA and immunoreactivity in the buccal mucosa and arm skin of normal and immunodeficiency virus (SHIV/SIV) infected rhesus macaques. It was found that mmuIFNLR1 has 93.1% amino acid sequence identity to that of humans; all the amino acid residues of mmuIFNLR1 signal peptide, transmembrane region, PxxLxF motif and those essential for ligand binding are identical to that of humans; 6 variants of mmuIFNLR1, including the ones corresponding to that of humans were detected; IFNLR1 immunoreactivity was localized in primarily the epithelia of buccal mucosa and arm skin; SHIV/SIV infection could affect the levels of mmuIFNLR1 mRNA and immunoreactivity. These data expanded our knowledge on mmuIFNLR1 and provided a scientific basis for rational use of rhesus macaques in studies of IFN-λ associated human diseases like AIDS. Future studies testing IFNLR1-targeting therapeutics in rhesus macaques were warranted.
RESUMEN
Interleukin-17A (IL-17A) and interleukin-17F (IL-17F) as two potent proinflammatory cytokines and the signature cytokines of Th17 cells play important roles in human autoimmune diseases, inflammation and host defenses. In this study, rhesus macaque IL-17A (rhIL-17A) and IL-17F (rhIL-17F) were cloned and expressed, and their biological activities and in vivo distribution were examined. The resulting data showed that both the rhIL-17A and rhIL-17F genes were consisted of three exons and two introns. RhIL-17A and rhIL-17F shared 96.8% and 93.9% amino acid sequence identity with human IL-17A (huIL-17A) and IL-17F (huIL-17F) respectively and the sequences also shared one N-glycosylation site and six conserved cysteine residues with huIL-17A and huIL-17F. IL-17A and IL-17F transcripts were highly expressed in lymphoid tissues and the intestinal tract of rhesus macaques. Functionally, recombinant rhIL-17A and rhIL-17F showed similar effect on Act1 levels and NF-κB phosphorylation compared with that of commercial human IL-17A and IL-17F. Moreover, the antibacterial proteins (such as ß-defensin 2, S100A8, S100A9, RegIIIα and Muc1) and the tight junction associated genes (including CLDN1, CLDN4, OCLN, and ZO1) expressed by Caco-2 cells were largely enhanced after treatment with rhIL-17A and rhIL-17F. Meanwhile, purified rhIL-17A and rhIL-17F could also induce the expression of IL-6 and TNF-α by THP-1 cells. These data indicated that rhesus macaque IL-17A and IL-17F are highly similar to that of humans in both structure and function. Studies on rhIL-17A/rhIL-17F are promising approach to contribute to the understanding of human IL-17A and IL-17F-related intestinal diseases.
Asunto(s)
Interleucina-17/genética , Interleucina-17/metabolismo , Animales , Células CACO-2 , Línea Celular , Clonación Molecular/métodos , Citocinas/genética , Citocinas/metabolismo , Humanos , Inflamación/genética , Inflamación/metabolismo , Macaca mulatta , FN-kappa B/genética , FN-kappa B/metabolismo , Células THP-1 , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
It is widely accepted that impairment of the intestinal epithelial barrier from HIV/AIDS contributes significantly to microbial translocation and systemic immune activation. Such factors present potential targets for novel treatments aimed toward a functional cure. However, the extracellular mechanisms of intestinal barrier repair are poorly understood. In the current study, we investigated the abilities of IL-17A and IL-17F to repair the damaged barrier caused by HIV-1 gp140 using Caco-2 monolayers. It was found that HIV-1 gp140 downregulated the expression of tight junction-associated genes and disrupted the barrier integrity of Caco-2 monolayers. However, IL-17A and IL-17F treatment reversed the HIV-1 gp140-induced barrier dysfunction by upregulating the expression of tight junction-associated genes, the combination of which resulted in a stronger induction of barrier repair. Furthermore, the effects of IL-17A and IL-17F were reduced by downregulation of Act1 with siRNA and inhibition of NF-κB and MAPK pathways with BAY11-7082 and U0126, respectively. These data indicated that the NF-κB and MAPK pathways are involved in the repair of barrier integrity mediated by IL-17A and IL-17F, and IL-17 pathways are potential targets for gut barrier restoration therapies during HIV/AIDS.
Asunto(s)
Infecciones por VIH/fisiopatología , Interleucina-17/farmacología , Mucosa Intestinal/efectos de los fármacos , Proteínas de Uniones Estrechas/genética , Uniones Estrechas/genética , Productos del Gen env del Virus de la Inmunodeficiencia Humana/toxicidad , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Células CACO-2 , Infecciones por VIH/tratamiento farmacológico , Humanos , Permeabilidad/efectos de los fármacos , Fosforilación/efectos de los fármacos , ARN Interferente Pequeño/genética , Transducción de Señal/efectos de los fármacos , Proteínas de Uniones Estrechas/metabolismo , Uniones Estrechas/metabolismo , Factor de Transcripción ReIA/antagonistas & inhibidores , Factor de Transcripción ReIA/metabolismo , Regulación hacia Arriba , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis) are frequently used in establishing animal models for human diseases. To determine the differences in gut microbiota between these species, rectal swabs from 20 rhesus macaques and 21 cynomolgus macaques were collected, and the microbial composition was examined by deep sequencing of the 16S rRNA gene. We found that the rectal microbiota of cynomolgus macaques exhibited significantly higher alpha diversity than that of rhesus macaques, although the observed number of operational taxonomic units (OTUs) was almost the same. The dominant taxa at both the phylum and genus levels were similar between the two species, although the relative abundances of these dominant taxa were significantly different between them. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) showed significant differences in the functional components between the microbiota of the two species, in particular the lipopolysaccharide (LPS) synthesis proteins. The above data indicated significant differences in microbial composition and function between these two closely related macaque species, which should be taken into consideration in the future selection of these animals for disease models.
Asunto(s)
Bacterias/genética , Microbioma Gastrointestinal , Macaca fascicularis/microbiología , Macaca mulatta/microbiología , Recto/microbiología , Animales , Bacterias/clasificación , Genoma Bacteriano , Genómica , Especificidad de la EspecieRESUMEN
Interleukin-22 (IL-22) is a potential therapeutic agent for diseases driven by epithelial injury. To characterize the IL-22 expressed by rhesus macaques, animals that are irreplaceable for human disease research, rhesus macaque IL-22 (rhIL-22) was cloned and expressed, and its biological activity and in vivo distribution were examined. It was found that the rhIL-22 gene consists of five introns and six exons, including a short non-coding exon starting 22 bp downstream of a putative TATA box. The amino acid sequence of rhIL-22 showed 95·5% identity to that of humans, and it shared two conserved disulphide bonds, three N-glycosylation sites and all the critical residues for binding to IL-22R1. High levels of IL-22 mRNA were observed in the liver, pancreas, lymphoid tissues and especially in the outer-body barriers such as the intestinal tract of rhesus macaques. Functionally, purified rhIL-22 has a similar but a little earlier effect on signal transducer and activator of transcription 3 phosphorylation at Tyr705 compared with that of commercial human IL-22. The expression of the antibacterial proteins ß-defensin-2, S100A8, S100A9, RegIIIα and Muc1 by HT-29 cells was largely upregulated after stimulation with rhIL-22. Recombinant rhIL-22 could also significantly promote the proliferation of human intestinal epithelial cells without affecting cell apoptosis. These data indicate that rhesus macaque IL-22 is highly similar to that of humans in both structure and function, and tests of therapeutic effects of human IL-22 on human diseases in rhesus macaques are warranted.
Asunto(s)
Interleucinas/genética , Mucosa Intestinal/fisiología , Hígado/fisiología , Macaca mulatta/genética , Páncreas/fisiología , Animales , Péptidos Catiónicos Antimicrobianos/metabolismo , Proliferación Celular , Clonación Molecular , Regulación de la Expresión Génica , Glicosilación , Células HT29 , Humanos , Macaca mulatta/inmunología , Factor de Transcripción STAT3/metabolismo , Transcriptoma , Interleucina-22RESUMEN
DC, NK lectin group receptor-1 (DNGR-1), also known as C-type lectin domain family 9 member A (CLEC9A), is a promising target for immunological therapeutics and vaccination against tumors and viruses. However, little is known about its property in rhesus macaques. In this study, we cloned rhesus macaque DNGR-1 cDNA, and found that its coding region could encode a 241-amino acid polypeptide with 91.7% sequence identity and similar antigenicity to that of humans. Both free and cell surface rhesus macaque DNGR-1 expressed in vitro could bind to apoptotic/dead cells induced by serum deprivation or freeze-thaw, and to pyroptotic cells stimulated with PMA and LPS. We also demonstrated that rhesus macaque DNGR-1 mRNA was present in all the examined tissues, with the highest in lymph nodes, spleen, blood, and thymus. The expression of DNGR-1 that is highly similar to that of humans warranted the usefulness of rhesus macaques in testing human therapeutics and vaccines targeting DNGR-1, especially those for HIV/AIDS.
Asunto(s)
Lectinas Tipo C/genética , Macaca mulatta/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , Clonación Molecular/métodos , ADN Complementario/genética , Células HEK293 , Células HeLa , Humanos , ARN Mensajero/genética , ConejosRESUMEN
Dendritic cell NK lectin Group Receptor-1 (DNGR-1), also known as C-type lectin domain family 9, member A (CLEC9A), is a member of C-type lectin receptor superfamily expressed primarily on dendritic cells (DC) that excel in cross-presentation of exogenous antigens. To find out whether and how it is affected in human immunodeficiency virus infections or acquired immunodeficiency syndromes (HIV/AIDS), DNGR-1 expression and DNGR-1-binding cells in simian/human immunodeficiency virus (SHIV) and simian immunodeficiency virus (SIV)-infected rhesus macaques and antiretroviral therapy (ART)-treated AIDS patients were examined by real-time RT-PCR, flow cytometry, and confocal microscopy. DNGR-1 expression was observed in both lymphoid and non-lymphoid tissues including gut-associated lymphoid tissues (GALT) of rhesus macaques. DNGR-1 mRNA levels were significantly reduced in the blood while significantly elevated in the GALT of SHIV/SIV-infected rhesus macaques. DNGR-1 transcription levels were also significantly reduced in the blood of ART-treated AIDS patients irrespective of viral status. White blood cells with exposed DNGR-1 ligands were significantly increased in ART-treated AIDS patients, while significantly decreased in SIV-infected rhesus macaques. These data indicate that DNGR-1 expression, and by extension, the function of cross-presentation of antigens associated with dead/damaged cells might be compromised in HIV/SIV infection, which might play a role in HIV/AIDS pathogenesis and should be taken into consideration in therapeutic AIDS vaccine development.
Asunto(s)
Vacunas contra el SIDA/inmunología , Células Dendríticas/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Células Asesinas Naturales/inmunología , Lectinas Tipo C/metabolismo , Receptores Mitogénicos/metabolismo , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Virus de la Inmunodeficiencia de los Simios/inmunología , Animales , Antirretrovirales/uso terapéutico , Células Cultivadas , Reactividad Cruzada , Células Dendríticas/virología , Regulación de la Expresión Génica , Infecciones por VIH/tratamiento farmacológico , Humanos , Lectinas Tipo C/genética , Activación de Linfocitos , Macaca mulatta , Receptores Mitogénicos/genética , Receptores de Neuroquinina-1/metabolismoRESUMEN
Polymeric immunoglobulin receptors(pIgR) are key participants in the formation and secretion of secretory IgA(S-IgA), which is critical for the prevention of microbial infection and colonization in the respiratory system. Although increased respiratory colonization and infections are common in HIV/AIDS, little is known about the expression of pIgR in the airway mucosa of these patients. To address this, the expression levels of pIgR in the tracheal mucosa and lungs of SHIV/SIV-infected rhesus macaques were examined by real-time RTPCR and confocal microscopy. We found that the levels of both PIGR mRNA and pIgR immunoreactivity were lower in the tracheal mucosa of SHIV/SIV-infected rhesus macaques than that in non-infected rhesus macaques, and the difference in pIgR immunoreactivity was statistically significant. IL-17A, which enhances pIgR expression, was also changed in the same direction as that of pIgR. In contrast to changes in the tracheal mucosa, pIgR and IL-17A levels were higher in the lungs of infected rhesus macaques. These results indicated abnormal pIgR expression in SHIV/SIV, and by extension HIV infections, which might partially result from IL-17A alterations and might contribute to the increased microbial colonization and infection related to pulmonary complications in HIV/AIDS.
Asunto(s)
Receptores de Inmunoglobulina Polimérica/metabolismo , Mucosa Respiratoria/metabolismo , Síndrome de Inmunodeficiencia Adquirida del Simio/metabolismo , Virus de la Inmunodeficiencia de los Simios , Tráquea/virología , Animales , Regulación de la Expresión Génica , Macaca mulatta , Receptores de Inmunoglobulina Polimérica/genética , Síndrome de Inmunodeficiencia Adquirida del Simio/virologíaRESUMEN
BACKGROUND: Mucosal barrier dysfunction might play a key role in HIV/AIDS, yet the early effects of HIV-1 on intestinal mucosal barrier, especially tight junctions (TJ) have not been well addressed. AIMS: To investigate the effects of acute HIV-1 infection on the expression of intestinal IL-17A and TJ-associated genes using an NHP-AIDS model. METHODS: TaqMan probe real-time RT-PCR methods were established and claudin-1, claudin-3, occludin and zonula occluden-1 (ZO-1) mRNA levels in the duodenal biopsies of rhesus macaques collected before and after rectal exposures to SHIV-SF162P4 were examined and compared with that of IL-17A, IL-6, TGF-ß, RORγt, T-bet, Foxp-3 and GATA-3. RESULTS: The mRNA levels of TJ-associated genes were statistically significantly reduced soon after viral exposures and the mRNA levels of claudin-1, occludin and ZO-1 in viral positive tissues (from Group I) were lower than that in viral negative tissues (from Group II) after viral exposure. IL-17A mRNA levels were also decreased and positively correlated with the mRNA levels of the TJ-associated genes after viral exposure or infection, although the levels of IL-6, TGF-ß and RORγt mRNA showed no statistical difference. The levels of GATA-3 mRNA in tissues collected before viral exposure were statistically different between Group I and Group II animals. The balance between T-bet and GATA-3 mRNA levels in Group II was markedly altered and statistically significantly different from that in Group I. CONCLUSIONS: Acute SHIV, and by extension HIV infection could affect the expression of TJ-associated genes, probably through IL-17A and other immune alterations.
Asunto(s)
Duodeno/metabolismo , Infecciones por VIH/metabolismo , Interleucina-17/metabolismo , Mucosa Intestinal/metabolismo , Síndrome de Inmunodeficiencia Adquirida del Simio/metabolismo , Proteínas de Uniones Estrechas/metabolismo , Animales , Células Cultivadas , Duodeno/citología , Duodeno/virología , VIH-1/genética , VIH-1/patogenicidad , Interleucina-17/genética , Mucosa Intestinal/virología , Macaca mulatta , ARN Mensajero/genética , ARN Mensajero/metabolismo , Virus de la Inmunodeficiencia de los Simios/genética , Virus de la Inmunodeficiencia de los Simios/patogenicidad , Proteínas de Uniones Estrechas/genética , Transcripción GenéticaAsunto(s)
Proteínas Fluorescentes Verdes/análisis , Ganglios Linfáticos Agregados/virología , Síndrome de Inmunodeficiencia Adquirida del Simio/patología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/aislamiento & purificación , Bazo/virología , Animales , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Macaca mulatta , Virus de la Inmunodeficiencia de los Simios/genéticaRESUMEN
BACKGROUND: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1), a member of the immunoglobulin superfamily, is essential for gut-specific homing of leukocytes; however, it has not been well characterized in rhesus macaques. AIMS: To obtain the complete nucleotide sequence of rhesus macaque MAdCAM-1 cDNA and determine its distribution in gut-associated lymphoid tissues (GALT) and its alteration in duodenal mucosa after simian/human immunodeficiency virus (SHIV) infection. METHODS: MAdCAM-1 cDNA was cloned from the colon mucosa of a rhesus macaque by 3'- and 5'-RACE. The distribution and abundance of MAdCAM-1 mRNA in the GALT were examined by nested and real-time RT-PCR. The alterations of MAdCAM-1 mRNA levels in SHIV-infected duodenal mucosa were determined by real-time RT-PCR. RESULTS: The nucleotide sequence of rhesus macaque MAdCAM-1 cDNA (1,503 bp nucleotides) including the 5'- and 3'-untranslated regions was obtained. The coding region (1,086 bp) showed 87.56% and the Ig-like domain 1, 2 and TM + cytoplasmic domains showed >93% nucleotide sequence identity to that of humans. Like humans, rhesus macaques lacked MAdCAM-1 IgA1-like domain, which could be a common feature for all primates appeared later during vertebrate evolution. Two species of MAdCAM-1 mRNA were detected and high-level transcripts were observed primarily in the GALT. The full-length MAdCAM-1 expressed in vitro could bind to human α4ß7. MAdCAM-1 mRNA levels were statistically significantly reduced in SHIV-infected duodenal mucosa. CONCLUSIONS: These data provided a basis for using rhesus macaques in pathological and therapeutic studies on leukocyte homing related diseases such as inflammatory bowel disease and HIV/AIDS.
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Clonación Molecular , Regulación de la Expresión Génica/inmunología , Inmunoglobulinas/metabolismo , Mucoproteínas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Ácidos Carboxílicos/metabolismo , ADN Complementario , Humanos , Inmunoglobulinas/genética , Tejido Linfoide , Macaca mulatta , Datos de Secuencia Molecular , Mucoproteínas/genética , Naftalenos/metabolismo , Filogenia , Unión ProteicaRESUMEN
The roles of the interactions between nervous, endocrine, and immune systems have been well established in human health and diseases. At present, little is known about the cellular bases for neural-endocrine-immune networks in the gastrointestinal mucosa. In the current study, duodenum, jejunum, ileum, cecum, colon, and rectum autopsies from 15 rhesus macaques and endoscopic duodenal biopsies from 12 rhesus macaques were collected, and the spatial relationships between the endocrine cells and immune cells in the intestinal mucosa were examined by transmission electron microscopy. Eight types of enteroendocrine cells similar to human enterochromaffin cells (EC), D1, G, I, K, L, N, and S cells were found to lie within a one-cell-size distance from immunocytes, in particular the eosinophils in the epithelia or lamina propria. Close apposition of large areas of plasma membranes between many types of enteroendocrine cells and immunocytes, especially between EC, K, S cells and eosinophils, were observed in the epithelia for the first time. These data indicate that complex interactions occur between diverse types of enteroendocrine cells and various immune cells through paracrine mechanisms or via mechanisms dependent on cell-to-cell contact; such interactions might play key roles in maintaining the gut mucosal barrier integrity of rhesus macaques.
Asunto(s)
Comunicación Celular , Células Enteroendocrinas/citología , Células Enteroendocrinas/inmunología , Mucosa Intestinal/citología , Mucosa Intestinal/inmunología , Linfocitos/citología , Macaca mulatta/inmunología , Animales , Membrana Celular/ultraestructura , Células Enteroendocrinas/ultraestructura , Eosinófilos/citología , Eosinófilos/ultraestructura , Humanos , Mucosa Intestinal/ultraestructura , Linfocitos/ultraestructuraRESUMEN
Genetic, epidemiological and experimental evidence suggest that the major histocompatibility complex (MHC) is critical in controlling human immunodeficiency virus (HIV) infection. The objectives of this study were to determine whether novel recombinant Mamu MHC constructs would elicit protection against rectal challenge with heterologous simian-human immunodeficiency virus (SHIV) strain SF162.P4 in rhesus macaques. Mamu class I and II gene products were linked together with HIV gp140, simian immunodeficiency virus (SIV) p27 and heat-shock protein 70 to dextran. The vaccine was administered to two groups, each consisting of nine macaques, either subcutaneously (SC), or rectally and boosted by SC immunization. The controls were untreated or adjuvant-treated animals. Repetitive rectal challenges with up to ten doses of SHIV SF162.P4 showed a significant decrease in the peak and sequential viral RNA concentrations, and three macaques remained uninfected, in the nine SC-immunized animals, compared with infection in all nine controls. Macaques immunized rectally followed by SC boosters showed a less significant decrease in both sequential and peak viral loads compared with the SC-immunized animals, and all were infected following rectal challenge with SHIV SF162.P4. Plasma and mucosal IgG and IgA antibodies to Mamu class I alleles and HIV gp120, as well as to RANTES (regulated upon activation, normal T-cell expressed, and secreted; CCR5) were increased, and showed significant inverse correlations with the peak viral load. These results suggested that allo-immunization with recombinant MHC constructs linked to HIV-SIV antigens merits further investigation in preventing HIV-1 infection.
Asunto(s)
Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Vacunas contra el SIDAS/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/prevención & control , Virus de la Inmunodeficiencia de los Simios/inmunología , Productos del Gen env del Virus de la Inmunodeficiencia Humana/inmunología , Administración Rectal , Animales , Anticuerpos Antivirales/sangre , Modelos Animales de Enfermedad , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Inyecciones Subcutáneas , Macaca mulatta , Vacunas contra el SIDAS/administración & dosificación , Vacunación/métodos , Carga ViralRESUMEN
CD1c+ myeloid dendritic cells (mDCs) in the peripheral blood of 30 SHIV-SF162p4 and SIVmac251 sequentially infected Chinese rhesus macaques were examined by flow cytometry to obtain further insight into mDC alterations in HIV/AIDS. The CD1c+ cells were found to be mononuclear leukocytes rather than granulocytes, and most of them expressed CD20. CD1c+mDCs (CD1c+CD20-) consisted of two morphological subsets: the granular and the large CD1c+mDCs. The expression of HLA-DR, CD86, and CD11b, but no CCR7, CD83 and CD123, together with their endocytotic capacity indicated that they were immature mDCs. Their frequency at weeks 10 and 12 post-infection was significantly higher than that of un-infected ones; the large CD1c+mDC level was significantly different between time points and almost absent from un-infected rhesus monkeys; significant correlations between CD1c+mDCs and plasma viral load levels were also observed. These data indicated a possible role for CD1c+mDCs in the pathophysiological process of SIV/HIV infection.
Asunto(s)
Antígenos CD1/inmunología , Células Dendríticas/inmunología , Infecciones por VIH/inmunología , Leucocitos Mononucleares/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Animales , Antígenos CD1/sangre , Antígenos CD20/sangre , Antígenos CD20/inmunología , Antígeno B7-2/sangre , Antígeno B7-2/inmunología , Antígeno CD11b/sangre , Antígeno CD11b/inmunología , China , Células Dendríticas/metabolismo , Endocitosis/inmunología , Citometría de Flujo , Infecciones por VIH/sangre , Infecciones por VIH/virología , Antígenos HLA-DR/sangre , Antígenos HLA-DR/inmunología , Humanos , Leucocitos Mononucleares/metabolismo , Leucocitos Mononucleares/virología , Macaca mulatta , Células Mieloides/inmunología , Células Mieloides/metabolismo , Síndrome de Inmunodeficiencia Adquirida del Simio/sangre , Síndrome de Inmunodeficiencia Adquirida del Simio/virologíaRESUMEN
Human immunodeficiency virus (HIV)-1 is mainly transmitted mucosally during sexual intercourse. We therefore evaluated the protective efficacy of a vaccine active at mucosal sites. Macaca mulatta monkeys were immunized via both the intramuscular and intranasal routes with an HIV-1 vaccine made of gp41-subunit antigens grafted on virosomes, a safe delivery carrier approved in humans with self-adjuvant properties. Six months after 13 vaginal challenges with simian-HIV (SHIV)-SF162P3, four out of five vaccinated animals remained virus-negative, and the fifth was only transiently infected. None of the five animals seroconverted to p27gag-SIV. In contrast, all 6 placebo-vaccinated animals became infected and seroconverted. All protected animals showed gp41-specific vaginal IgAs with HIV-1 transcytosis-blocking properties and vaginal IgGs with neutralizing and/or antibody-dependent cellular-cytotoxicity activities. In contrast, plasma IgGs totally lacked virus-neutralizing activity. The protection observed challenges the paradigm whereby circulating antiviral antibodies are required for protection against HIV-1 infection and may serve in designing a human vaccine against HIV-1-AIDS.
Asunto(s)
Vacunas contra el SIDA/administración & dosificación , Anticuerpos Anti-VIH/biosíntesis , Proteína gp41 de Envoltorio del VIH/inmunología , VIH-1/inmunología , Inmunización , Macaca mulatta/inmunología , Fragmentos de Péptidos/inmunología , Vagina/inmunología , Virosomas/inmunología , Vacunas contra el SIDA/inmunología , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Citotoxicidad Celular Dependiente de Anticuerpos , Sitios de Unión , Femenino , Anticuerpos Anti-VIH/inmunología , Proteína gp41 de Envoltorio del VIH/administración & dosificación , Infecciones por VIH/inmunología , Infecciones por VIH/prevención & control , Infecciones por VIH/transmisión , Seropositividad para VIH , Datos de Secuencia Molecular , Fragmentos de Péptidos/administración & dosificación , Transcitosis , Viremia/inmunología , Viremia/prevención & control , Viremia/transmisión , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/análisisRESUMEN
Serotonin transporter (SERT, 5-HTT) is a key element in the serotonergic system which is probably involved in the psychiatric disorders commonly observed in people living with HIV/AIDS. However, no information is available about the effects of HIV infection on SERT expression. In this study, a TaqMan real-time RT-PCR method was established, levels of SERT mRNA in the peripheral blood mononuclear cells (PBMCs) and various tissues from normal Chinese rhesus macaques, in PBMCs from 32 SHIV-sf162p4 infected rhesus macaques and from 8 rhesus macaques before and 7, 14, 21, 28 and 196 days after SHIV-sf162p4 infection, and in PBMCs before and after in vitro phytohemagglutinin (PHA) stimulation were examined. It was found that SERT mRNA was widely distributed in lymphoid tissues; the level of SERT mRNA was significantly reduced in PBMCs from SHIV infected rhesus macaques and in PBMCs stimulated with PHA. The most evident decrease (to about one-tenth) in SERT mRNA level was observed at day 7 after SHIV infection. Difference in PBMC SERT mRNA level between 5-HTTLPR genotypes was not statistically significant. These data indicated that, in addition to previously observed abnormality in serotonin metabolism, SERT expression might be affected in HIV/AIDS, which might be associated with depression and other psychiatric disorders in HIV/AIDS. Besides, this study provided a basis for quantitative analysis of SERT gene expression under effects of host and environmental factors, such as 5-HTTLPR genotypes, SERT targeting drugs or other infectious agents.
Asunto(s)
Infecciones por VIH/metabolismo , Monocitos/metabolismo , ARN Mensajero/biosíntesis , Proteínas de Transporte de Serotonina en la Membrana Plasmática/biosíntesis , Síndrome de Inmunodeficiencia Adquirida del Simio/metabolismo , Animales , Línea Celular , Cartilla de ADN , VIH-1 , Humanos , Macaca mulatta , Activación de Macrófagos/inmunología , Activación de Macrófagos/fisiología , Monocitos/efectos de los fármacos , Fitohemaglutininas , Polimorfismo Genético/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas de Transporte de Serotonina en la Membrana Plasmática/genética , Virus de la Inmunodeficiencia de los SimiosRESUMEN
BACKGROUND: The CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque. METHODS: A SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion. RESULTS: One SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection. CONCLUSIONS: We conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.
Asunto(s)
Genes env , VIH-1/genética , Virus de la Inmunodeficiencia de los Simios/genética , Animales , Quimera , VIH-1/fisiología , Humanos , Macaca mulatta , Provirus/genética , Receptores CCR5/fisiología , Virus de la Inmunodeficiencia de los Simios/fisiologíaRESUMEN
OBJECTIVE: To develop a model of SHIV-KB9/Chinese origin rhesus (Ch Rh) macaques for vaccine research and to compare the pathogenesis of SHIV-KB9 in Ch Rh macaques with that reported in Indian rhesus (Ind Rh) macaques. METHODS: Seven mamu-A*01 negative Ch Rh macaques were inoculated intravenously with 1-10000 MID50 of SHIV-KB9. The monkeys were monitored for viral load, CD4, CD8, SHIV-specific antibody and virus genetic variation. The results were compared with those previously observed in Ind Rh macaques. RESULTS: As compared to that observed in Ind Rh macaques, SHIV-KB9 in Ch Rh macaques displayed three identical disease progression patterns. However, the primary pattern was not identical between the two subspecies. The level of plasma viremia differed in SHIV-KB9-infected Ch Rh macaques which exhibited different outcomes from those in Ind Rh macaques. Generally, the values of viral load and the maintenance of CD4+ T cells were associated with humoral responses. Otherwise, the viral genetic distances (divergence, diversity) were larger in animals (M419, M425) with their CD4+ T cells profoundly depleted. CONCLUSION: The model of SHIV-KB9/Ch Rh macaques displays a relatively slow progression to AIDS compared with Ind Rh macaques, which may more accurately reflect the potential of candidate vaccines in humans.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida del Simio/patología , Animales , Relación CD4-CD8 , China , Progresión de la Enfermedad , India , Macaca mulatta , Especificidad de la Especie , Carga ViralRESUMEN
Rhesus macaques have long been used in animal models for various human diseases, the susceptibility and/or resistance to some of which have been associated with the major histocompatibilty complex (MHC). To gain insight into the MHC background and to facilitate the experimental use of Chinese rhesus macaques, the second exon of MhcMamu-DQB1 genes in 105 rhesus macaques were characterized by cloning and sequencing. A total of 37 MhcMamu-DQB1 alleles were identified, illustrating a marked allelic polymorphism at DQB1 in these monkeys. In addition to 10 alleles were novel sequences that had not been documented in earlier reports, at least 14 alleles reported in earlier studies were not detected in this study. Most of the sequences (73%) observed in this study belong to DQB1 06 (13 alleles) and DQB1 18 (14 alleles) lineages, and the rest (27%) belong to DQB1 15, DQB1 16 and DQB1 17 lineages. The most frequent allele detected among these monkeys was MhcMamu-DQB1 06111 (22%), followed by DQB1 1503 (19%); and most of the novel alleles were present at a frequency of less than 2.5%. As for individual animals, 24 of 105 (23%) were homozygous whereas 81 of 105 (77%) were heterozygous at the MhcMamu-DQB1 locus. These data indicated significant differences in MhcMamu-DQB1 allele distribution between the Chinese rhesus macaques and the previously reported rhesus macaques, which were mostly of Indian origin. This information will not only promote the understanding of rhesus macaque MHC diversity and polymorphism but will also facilitate the use of Chinese rhesus macaques in human disease studies, especially those that may be associated with HLA-DQB genes.
Asunto(s)
Genes MHC Clase II , Haplotipos , Macaca mulatta/genética , Alelos , Secuencia de Aminoácidos , Animales , Frecuencia de los Genes , Antígenos de Histocompatibilidad Clase II/química , Datos de Secuencia Molecular , Filogenia , Polimorfismo Genético , Alineación de SecuenciaRESUMEN
Available data on the normal levels of white blood cell populations in healthy rhesus macaques (Macaca mulatta) originated and living in China is scanty. To obtain such data, blood samples from 150 Chinese rhesus macaques were collected and the normal range of white blood cells and their subsets were analyzed according to age and sex by flow cytometry. CBC data showed that the count of total white blood cells and lymphocytes decreased with age. Phenotypic analysis of CD4 and CD8 expression on CD3+ T lymphocytes showed that the percentage of CD4+ T cells (51.4+/-9.6%), CD4-CD8- T cells (8.5+/-4.1%) and the ratio of CD4+ T to CD8+ T cells (1.26+/-0.55) decreased with age; and the percentage of CD8+ T cells (42.0+/-9.7%), CD4+CD8+ T cells (1.3+/-0.9%) and CD3+ lymphocytes (55.3+/-13.3%) increased with age. However, no statistically significant difference was observed between the male and female groups in most parameters in these monkeys except for the percentage of CD4+CD8+ T cells. This study provided basic information about blood cell count and T lymphocyte subsets in Chinese rhesus macaques. It may be useful for comparative studies using Indian and Chinese rhesus macaques.