RESUMO
Chimeric antigen receptor (CAR)-T cell therapy has become an important method for the treatment of hematological tumors. Lentiviruses are commonly used gene transfer vectors for preparing CAR-T cells, and the conditions for preparing CAR-T cells vary greatly. This study reported for the first time the influence of differences in infection temperature on the phenotype and function of produced CAR-T cells. Our results show that infection at 4 degrees produces the highest CAR-positive rate of T cells, infection at 37 degrees produces the fastest proliferation in CAR-T cells, and infection at 32 degrees produces CAR-T cells with the greatest proportion of naive cells and the lowest expression of immune checkpoints. Therefore, infection at 32 degrees is recommended to prepare CAR-T cells. CAR-T cells derived from infection at 32 degrees seem to have a balance between function and phenotype. Importantly, they have increased oncolytic ability. This research will help optimize the generation of CAR-T cells and improve the quality of CAR-T cell products.
Assuntos
Vetores Genéticos/genética , Neoplasias Hematológicas/terapia , Imunoterapia Adotiva/métodos , Infecções por Lentivirus/imunologia , Lentivirus/genética , Animais , Citotoxicidade Imunológica , Modelos Animais de Doenças , Células HEK293 , Xenoenxertos , Humanos , Masculino , Camundongos , Fenótipo , TemperaturaRESUMO
Mammals have developed clever adaptive and innate immune defense mechanisms to protect against invading bacterial and viral pathogens. Human innate immunity is continuously evolving to expand the repertoire of restriction factors and one such family of intrinsic restriction factors is the APOBEC3 (A3) family of cytidine deaminases. The coordinated expression of seven members of the A3 family of cytidine deaminases provides intrinsic immunity against numerous foreign infectious agents and protects the host from exogenous retroviruses and endogenous retroelements. Four members of the A3 proteins-A3G, A3F, A3H, and A3D-restrict HIV-1 in the absence of virion infectivity factor (Vif); their incorporation into progeny virions is a prerequisite for cytidine deaminase-dependent and -independent activities that inhibit viral replication in the host target cell. HIV-1 encodes Vif, an accessory protein that antagonizes A3 proteins by targeting them for polyubiquitination and subsequent proteasomal degradation in the virus producing cells. In this review, we summarize our current understanding of the role of human A3 proteins as barriers against HIV-1 infection, how Vif overcomes their antiviral activity, and highlight recent structural and functional insights into A3-mediated restriction of lentiviruses.
Assuntos
Desaminases APOBEC/fisiologia , Infecções por Lentivirus/imunologia , Lentivirus/imunologia , Desaminases APOBEC/química , Animais , Sequência de Bases , Humanos , Lentivirus/metabolismo , Infecções por Lentivirus/metabolismo , Estrutura Terciária de Proteína , Relação Estrutura-AtividadeRESUMO
RIG-I-like receptors (RLRs), as key cytoplasmic sensors of viral pathogen-associated molecular patterns, can recognise viral RNA and enhance the antiviral response. Some investigations have focused on the roles of RLRs in the innate immune response in grass carp, large yellow croaker, and rainbow trout. However, little is known about the function of RLRs in mandarinfish (Siniperca chuatsi), an important economic fish in Perciformes. Here, we functionally characterized the RLRs involved in the immune responses of mandarinfish (Siniperca chuatsi), by evaluating three RLRs, namely, RIG-I, MDA5, and LGP2. The results revealed that MDA5 and LGP2 were present in mandarinfish, whereas RIG-I was absent. The MDA5 and LGP2 cDNA sequences contained 2976 and 2046 bp and encoded 991 and 681 amino acids, respectively. Multiple sequence alignments showed that MDA5 and LGP2 of mandarinfish were clustered together with their homologs from other teleost fishes and shared high similarities with those from other vertebrates, and RIG-I of mandarinfish was absent. Moreover, quantitative real-time PCR (qPCR) analysis suggested that MDA5 and LGP2 were constitutively expressed in all tissues tested, and MDA5 mRNA expression was relatively high in the gill, and spleen, whereas LGP2 mRNA expression was high in the liver, gill, and head kidney. After stimulation with lipopolysaccharide or poly I:C, the expression of MDA5 and LGP2 was upregulated in spleen, gill and head kidney, but the pattern was not exactly the same, MDA5 transcripts generally increased and then declined with the prolonged infection, while LGP2 transcripts went up continuously, which showed that mandarinfish MDA5 and LGP2 may play independent roles in antiviral response. Besides, it is further revealed that the MDA5 could activate NF-κB and IRF3 to inducing the production of IFN-ß by constructing tet-on stable strain of 293T cell, however over-expression of LGP2 resulted in decreased NF-κB, IRF3 and IFN-ß production in cells challenged with LPS and polyI:C Taken together, our results demonstrated that MDA5 and LGP2, as a positive and negative regulator, respectively, played an important role in modulating antibacterial andantiviral immune responses though activating NF-κB and IRF3 in RLRs signaling of mandarinfish.
Assuntos
Proteínas de Peixes/metabolismo , Fatores Reguladores de Interferon/metabolismo , Helicase IFIH1 Induzida por Interferon/metabolismo , NF-kappa B/metabolismo , Perciformes/metabolismo , RNA Helicases/metabolismo , Animais , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Proteínas de Peixes/genética , Regulação da Expressão Gênica , Células HEK293 , Humanos , Imunidade Inata , Helicase IFIH1 Induzida por Interferon/genética , Lentivirus , Infecções por Lentivirus/imunologia , Lipopolissacarídeos/farmacologia , Poli I-C/imunologia , RNA Helicases/genética , Alinhamento de Sequência , Análise de Sequência de Proteína , Transdução de SinaisRESUMO
Immunosuppression is a hallmark of tumor progression, and treatments that inhibit or deplete monocytic myeloid-derived suppressive cells could promote anti-tumor immunity. c-FLIP is a central regulator of caspase-8-mediated apoptosis and necroptosis. Here we show that low-dose cytotoxic chemotherapy agents cause apoptosis linked to c-FLIP down-regulation selectively in monocytes. Enforced expression of c-FLIP or viral FLIP rescues monocytes from cytotoxicity and concurrently induces potent immunosuppressive activity, in T cell cultures and in vivo models of tumor progression and immunotherapy. FLIP-transduced human blood monocytes can suppress graft versus host disease. Neither expression of FLIP in granulocytes nor expression of other anti-apoptotic genes in monocytes conferred immunosuppression, suggesting that FLIP effects on immunosuppression are specific to monocytic lineage and distinct from death inhibition. Mechanistically, FLIP controls a broad transcriptional program, partially by NF-κB activation. Therefore, modulation of FLIP in monocytes offers a means to elicit or block immunosuppressive myeloid cells.
Assuntos
Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/imunologia , Infecções por Lentivirus/imunologia , Monócitos/imunologia , NF-kappa B/imunologia , Apoptose , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Células Cultivadas , Humanos , Terapia de Imunossupressão , Lentivirus/fisiologia , Infecções por Lentivirus/genética , Infecções por Lentivirus/fisiopatologia , Infecções por Lentivirus/virologia , Células Mieloides/imunologia , NF-kappa B/genéticaRESUMO
Lentiviruses are infectious agents of a number of animal species, including sheep, goats, horses, monkeys, cows, and cats, in addition to humans. As in the human case, the host immune response fails to control the establishment of chronic persistent infection that finally leads to a specific disease development. Despite intensive research on the development of lentivirus vaccines, it is still not clear which immune responses can protect against infection. Viral mutations resulting in escape from T-cell or antibody-mediated responses are the basis of the immune failure to control the infection. The innate immune response provides the first line of defense against viral infections in an antigen-independent manner. Antiviral innate responses are conducted by dendritic cells, macrophages, and natural killer cells, often targeted by lentiviruses, and intrinsic antiviral mechanisms exerted by all cells. Intrinsic responses depend on the recognition of the viral pathogen-associated molecular patterns (PAMPs) by pathogen recognition receptors (PRRs), and the signaling cascades leading to an antiviral state by inducing the expression of antiviral proteins, including restriction factors. This review describes the latest advances on innate immunity related to the infection by animal lentiviruses, centered on small ruminant lentiviruses (SRLV), equine infectious anemia virus (EIAV), and feline (FIV) and bovine immunodeficiency viruses (BIV), specifically focusing on the antiviral role of the major restriction factors described thus far.
Assuntos
Regulação da Expressão Gênica/imunologia , Imunidade Inata , Fatores Reguladores de Interferon/imunologia , Infecções por Lentivirus/imunologia , Receptores de Reconhecimento de Padrão/imunologia , Animais , Gatos , Bovinos , Células Dendríticas/imunologia , Células Dendríticas/virologia , Cabras , Cavalos , Vírus da Imunodeficiência Bovina/imunologia , Vírus da Imunodeficiência Bovina/patogenicidade , Vírus da Imunodeficiência Felina/imunologia , Vírus da Imunodeficiência Felina/patogenicidade , Vírus da Anemia Infecciosa Equina/imunologia , Vírus da Anemia Infecciosa Equina/patogenicidade , Fatores Reguladores de Interferon/genética , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/virologia , Infecções por Lentivirus/genética , Infecções por Lentivirus/virologia , Macrófagos/imunologia , Macrófagos/virologia , Moléculas com Motivos Associados a Patógenos/imunologia , Receptores de Reconhecimento de Padrão/genética , Ovinos , Linfócitos T/imunologia , Linfócitos T/virologiaRESUMO
CD8⺠T cells are critical for controlling viremia during human immunodeficiency virus (HIV) infection. These cells produce cytolytic factors and antiviral cytokines that eliminate virally- infected cells. During the chronic phase of HIV infection, CD8⺠T cells progressively lose their proliferative capacity and antiviral functions. These dysfunctional cells are unable to clear the productively infected and reactivated cells, representing a roadblock in HIV cure. Therefore, mechanisms to understand CD8⺠T cell dysfunction and strategies to boost CD8⺠T cell function need to be investigated. Using the feline immunodeficiency virus (FIV) model for lentiviral persistence, we have demonstrated that CD8⺠T cells exhibit epigenetic changes such as DNA demethylation during the course of infection as compared to uninfected cats. We have also demonstrated that lentivirus-activated CD4âºCD25⺠T regulatory cells induce forkhead box P3 (Foxp3) expression in virus-specific CD8⺠T cell targets, which binds the interleukin (IL)-2, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ promoters in these CD8⺠T cells. Finally, we have reported that epigenetic modulation reduces Foxp3 binding to these promoter regions. This review compares and contrasts our current understanding of CD8⺠T cell epigenetics and mechanisms of lymphocyte suppression during the course of lentiviral infection for two animal models, FIV and simian immunodeficiency virus (SIV).
Assuntos
Linfócitos T CD8-Positivos/imunologia , Epigênese Genética , Vírus da Imunodeficiência Felina , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/veterinária , Vírus da Imunodeficiência Símia , Animais , Gatos , Citocinas/imunologia , Infecções por HIV/imunologia , Humanos , Ativação Linfocitária , Linfócitos T Reguladores/imunologia , Viremia/veterináriaRESUMO
(1) Background: Small ruminant lentiviruses (SRLV) persist in infected goats that mount a strong humoral immune response characterized by low neutralizing titers. In this study, we characterized the antibody response to SU5, a variable, immunodominant epitope of the envelope glycoprotein of SRLV. We tested the working hypothesis that the variability of SU5 reflects escape from neutralizing antibody. (2) Methods: Affinity purified anti-SU5 antibody were tested for their neutralizing activity to the homologous lentivirus. Virus culture supernatant—in native form or following sonication and filtration—was used to test the ability of free envelope glycoproteins to compete for binding in a SU5-peptide-ELISA. (3) Results: Anti-SU5 antibodies are not neutralizing, strongly suggesting that they do not bind intact viral particles. In contrast, shed envelope glycoproteins efficiently compete for binding in a SU5-ELISA, providing convincing evidence that the SU5 epitope is exposed only on shed envelope glycoproteins. (4) Conclusions: Our results show that the antibody engaging SU5 is not neutralizing and does not appear to bind to SU expressed at the surface of virus particles. We propose that SU5 is a potential decoy epitope exposed on shaded envelope glycoproteins, luring the humoral immune response in committing an original antigenic sin to a functionally irrelevant epitope.
Assuntos
Anticorpos Neutralizantes/imunologia , Antígenos Virais/imunologia , Vírus da Artrite-Encefalite Caprina/imunologia , Epitopos Imunodominantes/imunologia , Proteínas do Envelope Viral/imunologia , Animais , Anticorpos Antivirais/imunologia , Antígenos Virais/genética , Ensaio de Imunoadsorção Enzimática , Doenças das Cabras/imunologia , Doenças das Cabras/virologia , Cabras/imunologia , Cabras/virologia , Epitopos Imunodominantes/genética , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/veterinária , Testes de Neutralização , Peptídeos/imunologia , Proteínas do Envelope Viral/genéticaRESUMO
The exact influence of caprine arthritis encephalitis virus (CAEV) infection on blood and milk polymorphonuclear leukocytes (PMNLs) and monocyte/macrophages of goats remains unclear. Thus, the present study sought to explore the blood and milk PMNL and monocyte/macrophage functions in naturally CAEV-infected goats. The present study used 18 healthy Saanen goats that were segregated according to sera test outcomes into serologically CAEV negative (n=8; 14 halves) and positive (n=10; 14 halves) groups. All milk samples from mammary halves with milk bacteriologically positive outcomes, somatic cell count ≥2×106cellsmL-1, and abnormal secretions in the strip cup test were excluded. We evaluated the percentage of blood and milk PMNLs and monocyte/macrophages, the viability of PMNLs and monocyte/macrophages, the levels of intracellular reactive oxygen species (ROS) and the nonopsonized phagocytosis of Staphylococcus aureus and Escherichia coli by flow cytometry. In the present study, a higher percentage of milk macrophages (CD14+) and milk polymorphonuclear leukocytes undergoing late apoptosis or necrosis (Annexin-V+/Propidium iodide+) was observed in CAEV-infected goats; we did not find any further alterations in blood and milk PMNL and monocyte/macrophage functions. Thus, regarding our results, the goats naturally infected with CAEV did not reveal pronounced dysfunctions in blood and milk polymorphonuclear leukocytes and monocytes/macrophages.
Assuntos
Vírus da Artrite-Encefalite Caprina/imunologia , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Macrófagos/imunologia , Monócitos/imunologia , Neutrófilos/imunologia , Animais , Feminino , Doenças das Cabras/sangue , Doenças das Cabras/imunologia , Cabras/sangue , Cabras/imunologia , Cabras/virologia , Infecções por Lentivirus/sangue , Infecções por Lentivirus/imunologia , Leite/citologia , Leite/imunologia , Leite/virologiaRESUMO
SIV DNA can be detected in lymphoid tissue-resident macrophages of chronically SIV-infected Asian macaques. These macrophages also contain evidence of recently phagocytosed SIV-infected CD4+ T cells. Here, we examine whether these macrophages contain replication-competent virus, whether viral DNA can be detected in tissue-resident macrophages from antiretroviral (ARV) therapy-treated animals and humans, and how the viral sequences amplified from macrophages and contemporaneous CD4+ T cells compare. In ARV-naive animals, we find that lymphoid tissue-resident macrophages contain replication-competent virus if they also contain viral DNA in ARV-naive Asian macaques. The genetic sequence of the virus within these macrophages is similar to those within CD4+ T cells from the same anatomic sites. In ARV-treated animals, we find that viral DNA can be amplified from lymphoid tissue-resident macrophages of SIV-infected Asian macaques that were treated with ARVs for at least 5 months, but we could not detect replication-competent virus from macrophages of animals treated with ARVs. Finally, we could not detect viral DNA in alveolar macrophages from HIV-infected individuals who received ARVs for 3 years and had undetectable viral loads. These data demonstrate that macrophages can contain replication-competent virus, but may not represent a significant reservoir for HIV in vivo.
Assuntos
DNA Viral/metabolismo , Infecções por HIV/virologia , HIV-1/genética , Macrófagos/virologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/genética , Replicação Viral , Animais , Antirretrovirais/uso terapêutico , Terapia Antirretroviral de Alta Atividade , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/virologia , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , HIV-1/fisiologia , Humanos , Infecções por Lentivirus/tratamento farmacológico , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Tecido Linfoide/citologia , Macaca , Macaca mulatta , Macaca nemestrina , Macrófagos/imunologia , Macrófagos/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/tratamento farmacológico , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/fisiologia , Carga ViralRESUMO
The conditioning regimen used as part of the Berlin patient's hematopoietic cell transplant likely contributed to his eradication of HIV infection. We studied the impact of conditioning in simian-human immunodeficiency virus-infected (SHIV-infected) macaques suppressed by combination antiretroviral therapy (cART). The conditioning regimen resulted in a dramatic, but incomplete depletion of CD4+ and CD8+ T cells and CD20+ B cells, increased T cell activation and exhaustion, and a significant loss of SHIV-specific Abs. The disrupted T cell homeostasis and markers of microbial translocation positively correlated with an increased viral rebound after cART interruption. Quantitative viral outgrowth and Tat/rev-induced limiting dilution assays showed that the size of the latent SHIV reservoir did not correlate with viral rebound. These findings identify perturbations of the immune system as a mechanism for the failure of autologous transplantation to eradicate HIV. Thus, transplantation strategies may be improved by incorporating immune modulators to prevent disrupted homeostasis, and gene therapy to protect transplanted cells.
Assuntos
Linfócitos T CD4-Positivos/efeitos da radiação , Linfócitos T CD8-Positivos/efeitos da radiação , Infecções por HIV/imunologia , HIV-1/efeitos da radiação , Transplante de Células-Tronco Hematopoéticas/métodos , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/efeitos da radiação , Condicionamento Pré-Transplante/métodos , Irradiação Corporal Total , Animais , Antirretrovirais/uso terapêutico , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Infecções por HIV/tratamento farmacológico , Homeostase/efeitos da radiação , Infecções por Lentivirus/tratamento farmacológico , Infecções por Lentivirus/imunologia , Macaca nemestrina , Síndrome de Imunodeficiência Adquirida dos Símios/tratamento farmacológico , Transplante Autólogo , Carga Viral/efeitos da radiaçãoRESUMO
The virus-host relationship in simian immunodeficiency virus (SIV) infected chimpanzees is thought to be different from that found in other SIV infected African primates. However, studies of captive SIVcpz infected chimpanzees are limited. Previously, the natural SIVcpz infection of one chimpanzee, and the experimental infection of six chimpanzees was reported, with limited follow-up. Here, we present a long-term study of these seven animals, with a retrospective re-examination of the early stages of infection. The only clinical signs consistent with AIDS or AIDS associated disease was thrombocytopenia in two cases, associated with the development of anti-platelet antibodies. However, compared to uninfected and HIV-1 infected animals, SIVcpz infected animals had significantly lower levels of peripheral blood CD4+ T-cells. Despite this, levels of T-cell activation in chronic infection were not significantly elevated. In addition, while plasma levels of ß2 microglobulin, neopterin and soluble TNF-related apoptosis inducing ligand (sTRAIL) were elevated in acute infection, these markers returned to near-normal levels in chronic infection, reminiscent of immune activation patterns in 'natural host' species. Furthermore, plasma soluble CD14 was not elevated in chronic infection. However, examination of the secondary lymphoid environment revealed persistent changes to the lymphoid structure, including follicular hyperplasia in SIVcpz infected animals. In addition, both SIV and HIV-1 infected chimpanzees showed increased levels of deposition of collagen and increased levels of Mx1 expression in the T-cell zones of the lymph node. The outcome of SIVcpz infection of captive chimpanzees therefore shares features of both non-pathogenic and pathogenic lentivirus infections.
Assuntos
Doenças dos Símios Antropoides/virologia , HIV-1/fisiologia , Infecções por Lentivirus/veterinária , Lentivirus de Primatas/fisiologia , Pan troglodytes , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/fisiologia , Animais , Doenças dos Símios Antropoides/imunologia , Doenças dos Símios Antropoides/patologia , Doenças dos Símios Antropoides/fisiopatologia , Doenças Autoimunes/etiologia , Doenças Autoimunes/veterinária , Biomarcadores/sangue , Contagem de Linfócito CD4 , Feminino , HIV-1/imunologia , HIV-1/isolamento & purificação , Hiperplasia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/fisiopatologia , Infecções por Lentivirus/virologia , Lentivirus de Primatas/imunologia , Lentivirus de Primatas/isolamento & purificação , Linfonodos/imunologia , Linfonodos/metabolismo , Linfonodos/patologia , Linfonodos/virologia , Masculino , Proteínas de Resistência a Myxovirus/metabolismo , Neopterina/sangue , Fragmentos de Peptídeos/sangue , Fragmentos de Peptídeos/química , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/sangue , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/química , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/patologia , Síndrome de Imunodeficiência Adquirida dos Símios/fisiopatologia , Vírus da Imunodeficiência Símia/imunologia , Vírus da Imunodeficiência Símia/isolamento & purificação , Trombocitopenia/etiologia , Trombocitopenia/veterinária , Carga Viral , Microglobulina beta-2/sangueRESUMO
Large granular lymphocytes (LGLs) have only been anecdotally reported in HIV infection. We previously reported an LGL lymphocytosis in FIV-infected cats associated with a rise in FIV proviral loads and a marked neutropenia that persisted during chronic infection. Extensive immunophenotyping of peripheral blood mononuclear cells in cats chronically infected with FIV were identified LGLs as CD8lo(+)FAS(+); this cell population expanded commensurate with viral load. CD8lo(+)FAS(+) cells expressed similar levels of interferon-γ compared to CD8lo(+)FAS(+) cells from FIV-naive control animals, yet CD3É expression, which was increased on total CD8(+) T cells in FIV-infected cats, was decreased on CD8lo(+)FAS(+) cells. Down-modulation of CD3 expression was reversed after culturing PBMC for 3 days in culture with ConA/IL-2. We identified CD8lo(+)FAS(+) LGLs to be polyclonal T cells lacking CD56 expression. Blood smears from HIV-infected individuals and SIVmac239-infected rhesus macaques revealed increased LGLs compared to HIV/SIV negative counterparts. In humans, there was no correlation with viral load or treatment and in macaques the LGLs arose in acute SIV infection with increases in viremia. This is the first report describing and partially characterizing LGL lymphocytosis in association with lentiviral infections in three different species.
Assuntos
Infecções por Lentivirus/veterinária , Linfócitos/imunologia , Animais , Complexo CD3/sangue , Antígeno CD56/sangue , Linfócitos T CD8-Positivos/imunologia , Doenças do Gato/sangue , Doenças do Gato/imunologia , Gatos , Síndrome de Imunodeficiência Adquirida Felina/sangue , Síndrome de Imunodeficiência Adquirida Felina/imunologia , Infecções por HIV/sangue , Infecções por HIV/imunologia , Humanos , Imunofenotipagem , Interferon gama/sangue , Infecções por Lentivirus/sangue , Infecções por Lentivirus/imunologia , Linfócitos/classificação , Linfócitos/patologia , Macaca mulatta , Síndrome de Imunodeficiência Adquirida dos Símios/sangue , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Especificidade da Espécie , Receptor fas/sangueRESUMO
BACKGROUND: Meningoencephalitis is one of the most common disorders of the central nervous system (CNS) worldwide. Viral meningoencephalitis differs from bacterial meningitis in several aspects. In some developing countries, bacterial meningitis has appropriate clinical management and chemotherapy is available. Virus-associated and virus not detected meningoencephalitis are treatable, however, they may cause death in a few cases. The knowledge of how mediators of inflammation can induce disease would contribute for the design of affordable therapeutic strategies, as well as to the diagnosis of virus not detected and viral meningoencephalitis. Cytokine-induced inflammation to CNS requires several factors that are not fully understood yet. METHODS: Considering this, several cytokines were measured in the cerebrospinal fluid (CSF) of patients with undiagnosed and viral meningoencephalitis, and these were correlated with cellularity in the CSF. RESULTS: The results demonstrate that an altered biochemical profile alongside increased cellularity in the cerebrospinal fluid is a feature of patients with meningoencephalitis that are not associated with the detection of virus in the CNS (P < 0.05). Moreover, HIV-positive patients (n = 10) that evolve with meningoencephalitis display a distinct biochemical/cytological profile (P < 0.05) in the cerebrospinal fluid. Meningoencephalitis brings about a prominent intrathecal cytokine storm regardless of the detection of virus as presumable etiological agent. In the case of Enterovirus infection (n = 13), meningoencephalitis elicits robust intrathecal pro-inflammatory cytokine pattern and elevated cellularity when compared to herpesvirus (n = 15) and Arbovirus (n = 5) viral infections (P < 0.05). CONCLUSION: Differences in the cytokine profile of the CSF may be unique if distinct, viral or presumably non-viral pathways initially trigger the inflammatory response in the CNS.
Assuntos
Infecções por Arbovirus/líquido cefalorraquidiano , Citocinas/líquido cefalorraquidiano , Infecções por Enterovirus/líquido cefalorraquidiano , Infecções por HIV/líquido cefalorraquidiano , Infecções por Herpesviridae/líquido cefalorraquidiano , Infecções por Lentivirus/líquido cefalorraquidiano , Meningoencefalite/líquido cefalorraquidiano , Infecções por Arbovirus/diagnóstico , Infecções por Arbovirus/imunologia , Viroses do Sistema Nervoso Central/líquido cefalorraquidiano , Viroses do Sistema Nervoso Central/diagnóstico , Viroses do Sistema Nervoso Central/imunologia , Coinfecção/líquido cefalorraquidiano , Coinfecção/imunologia , Estudos Transversais , Citocinas/imunologia , DNA Viral/líquido cefalorraquidiano , Infecções por Enterovirus/diagnóstico , Infecções por Enterovirus/imunologia , Infecções por HIV/diagnóstico , Infecções por HIV/imunologia , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/imunologia , Humanos , Inflamação , Interferon gama/líquido cefalorraquidiano , Interferon gama/imunologia , Interleucina-10/líquido cefalorraquidiano , Interleucina-10/imunologia , Interleucina-12/líquido cefalorraquidiano , Interleucina-12/imunologia , Interleucina-17/líquido cefalorraquidiano , Interleucina-17/imunologia , Interleucina-6/líquido cefalorraquidiano , Interleucina-6/imunologia , Infecções por Lentivirus/imunologia , Meningoencefalite/diagnóstico , Meningoencefalite/imunologia , RNA Viral/líquido cefalorraquidiano , Fator de Necrose Tumoral alfa/líquido cefalorraquidiano , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Neutralizing antibodies (NAbs) are believed to comprise an essential component of the protective immune response induced by vaccines against feline immunodeficiency virus (FIV) and human immunodeficiency virus (HIV) infections. However, relatively little is known about the role of NAbs in controlling FIV infection and subsequent disease progression. Here, we present studies where we examined the neutralization of HIV-luciferase pseudotypes bearing homologous and heterologous FIV envelope proteins (n = 278) by sequential plasma samples collected at 6 month intervals from naturally infected cats (n = 38) over a period of 18 months. We evaluated the breadth of the NAb response against non-recombinant homologous and heterologous clade A and clade B viral variants, as well as recombinants, and assessed the results, testing for evidence of an association between the potency of the NAb response and the duration of infection, CD4(+) T lymphocyte numbers, health status and survival times of the infected cats. Neutralization profiles varied significantly between FIV-infected cats and strong autologous neutralization, assessed using luciferase-based in vitro assays, did not correlate with the clinical outcome. No association was observed between strong NAb responses and either improved health status or increased survival time of infected animals, implying that other protective mechanisms were likely to be involved. Similarly, no correlation was observed between the development of autologous NAbs and the duration of infection. Furthermore, cross-neutralizing antibodies were evident in only a small proportion (13 %) of cats.
Assuntos
Anticorpos Neutralizantes/fisiologia , Especificidade de Anticorpos , Vírus da Imunodeficiência Felina/imunologia , Infecções por Lentivirus/veterinária , Animais , Linfócitos T CD4-Positivos , Gatos , Clonagem Molecular , Feminino , Regulação Viral da Expressão Gênica/fisiologia , Produtos do Gene env/genética , Produtos do Gene env/metabolismo , Células HEK293 , Humanos , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Masculino , Testes de NeutralizaçãoRESUMO
The feline immunodeficiency virus (FIV) shares genomic organization, receptor usage, lymphocyte tropism, and induction of immunodeficiency and increased susceptibility to cancer with the human immunodeficiency virus (HIV). Global distribution, marked heterogeneity and variable host adaptation are also properties of both viruses. These features render the FIV-cat model suitable to explore many aspects of lentivirus-host interaction and adaptation, and to explore treatment and prevention of infection. Examples of fundamental discoveries that have emerged from study in the FIV-cat model concern two-receptor entrance strategies that target memory T-lymphocytes, host factors that restrict retroviral infection, viral strategies for replication in non-dividing cells, and identification of correlates of immunity to the virus. This article provides a brief overview of strengths and limitations of the FIV-cat model for comparative biology and medicine.
Assuntos
Doenças do Gato/virologia , Modelos Animais de Doenças , Infecções por HIV/imunologia , HIV/fisiologia , Vírus da Imunodeficiência Felina/fisiologia , Infecções por Lentivirus/veterinária , Animais , Gatos , HIV/imunologia , Infecções por HIV/virologia , Humanos , Vírus da Imunodeficiência Felina/imunologia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Replicação ViralRESUMO
Th2-inducing pathological conditions such as parasitic diseases increase susceptibility to viral infections through yet unclear mechanisms. We have previously reported that IL-4, a pivotal Th2 cytokine, suppresses the response of murine bone-marrow-derived conventional dendritic cells (cDCs) and splenic DCs to Type I interferons (IFNs). Here, we analyzed cDC responses to TLR7 and TLR9 ligands, R848 and CpGs, respectively. We found that IL-4 suppressed the gene expression of IFNß and IFN-responsive genes (IRGs) upon TLR7 and TLR9 stimulation. IL-4 also inhibited IFN-dependent MHC Class I expression and amplification of IFN signaling pathways triggered upon TLR stimulation, as indicated by the suppression of IRF7 and STAT2. Moreover, IL-4 suppressed TLR7- and TLR9-induced cDC production of pro-inflammatory cytokines such as TNFα, IL-12p70 and IL-6 by inhibiting IFN-dependent and NFκB-dependent responses. IL-4 similarly suppressed TLR responses in splenic DCs. IL-4 inhibition of IRGs and pro-inflammatory cytokine production upon TLR7 and TLR9 stimulation was STAT6-dependent, since DCs from STAT6-KO mice were resistant to the IL-4 suppression. Analysis of SOCS molecules (SOCS1, -2 and -3) showed that IL-4 induces SOCS1 and SOCS2 in a STAT6 dependent manner and suggest that IL-4 suppression could be mediated by SOCS molecules, in particular SOCS2. IL-4 also decreased the IFN response and increased permissiveness to viral infection of cDCs exposed to a HIV-based lentivirus. Our results indicate that IL-4 modulates and counteracts pro-inflammatory stimulation induced by TLR7 and TLR9 and it may negatively affect responses against viruses and intracellular parasites.
Assuntos
Células Dendríticas/imunologia , Interleucina-4/fisiologia , Infecções por Lentivirus/imunologia , Glicoproteínas de Membrana/metabolismo , Receptor 7 Toll-Like/metabolismo , Receptor Toll-Like 9/metabolismo , Animais , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Células Dendríticas/virologia , Feminino , Imidazóis/farmacologia , Imunidade Inata , Fator Regulador 7 de Interferon/genética , Fator Regulador 7 de Interferon/metabolismo , Interleucina-6/metabolismo , Glicoproteínas de Membrana/agonistas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT2/genética , Fator de Transcrição STAT2/metabolismo , Proteína 1 Supressora da Sinalização de Citocina , Proteínas Supressoras da Sinalização de Citocina/genética , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Receptor 7 Toll-Like/agonistas , Ativação Transcricional/imunologia , Ubiquitinas/genética , Ubiquitinas/metabolismoRESUMO
Using the feline immunodeficiency virus (FIV) model for AIDS lentivirus infection, we previously demonstrated that Treg cells from FIV-infected cats up-regulate membrane-associated tumor growth factor beta (mTGF-ß) during the course of infection and that activated T lymphocytes up-regulate TGF-ß receptor II (TGF-ßRII) during the course of infection. Furthermore, we have demonstrated that autologous coculture of Tregs with Th cells from FIV-infected cats leads to suppression of interleukin (IL)-2 production and loss of proliferation in a TGF-ß-dependent fashion. Nuclear factor of activated T cells (NFAT) 2 has been identified as integral to effector Th cell maturation and function by promoting IL-2 transcription. Therefore, we questioned whether NFAT2 expression might be altered by TGF-ß signaling. Feline NFAT2 exon sequences were identified based upon sequence homology to human and murine NFAT2. Following stimulation, IL-2 and NFAT2 mRNA levels were similarly increased in both FIV(-) and FIV(+) cats. Activated CD4(+)CD25(-) cells from both FIV(-) and FIV(+) cats cocultured with autologous CD4(+)CD25(+) cells or treated with TGF-ß demonstrated decreased IL-2 production; however, NFAT2 mRNA levels were unaffected. Although NFAT2 mRNA levels were unaffected, chromatin immunoprecipitation (ChIP) for NFAT2 indicated decreased NFAT2 binding at the IL-2 promoter in suppressed Th cells. These data suggest that TGF-ß-mediated Treg cell suppression of IL-2 transcription is modulated through alterations in NFAT2 binding to the IL-2 promoter.
Assuntos
Vírus da Imunodeficiência Felina/imunologia , Interleucina-2/genética , Fatores de Transcrição NFATC/genética , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Reguladores/imunologia , Sequência de Aminoácidos , Animais , Gatos , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Interleucina-2/biossíntese , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/patologia , Infecções por Lentivirus/virologia , Linfonodos/citologia , Linfonodos/virologia , Ativação Linfocitária/imunologia , Dados de Sequência Molecular , Fatores de Transcrição NFATC/antagonistas & inibidores , Regiões Promotoras Genéticas/imunologia , Ligação Proteica/imunologia , RNA Mensageiro/biossíntese , Receptores de Fatores de Crescimento Transformadores beta/biossíntese , Alinhamento de Sequência , Linfócitos T Reguladores/virologia , Fator de Crescimento Transformador beta/biossíntese , Fator de Crescimento Transformador beta/farmacologia , Regulação para CimaRESUMO
Cat scratch disease is a zoonosis caused by Bartonella species, transmitted to humans through scratches or bites from infected cats and via direct contact with infected feces. Sporotrichosis, caused by the fungal complex Sporothrix, is transmitted by traumatic inoculation of the fungus. Cats are important in zoonotic transmission. Serum samples from 112 domestic cats with sporotrichosis and 77 samples from healthy cats were analyzed by indirect immunofluorescence assay (IFA), using the commercial kit Bartonella henselae IFA IgG (Bion). The presence of antibodies against feline leukemia virus (FeLV) and of feline immunodeficiency virus (FIV) core antigens was detected using the commercial kit Snap Combo FIV-FeLV (Idexx). The group of animals with sporotrichosis contained 93 males with a median age of 22 months, eight (7.1%) of which were positive for FIV and 15 (13.4%) for FeLV. The group of animals without sporotrichosis contained 36 males with a median age 48 months, 10 (13.0%) of which were positive for FIV and eight (10.4%) for FeLV. Of the 112 cats with sporotrichosis and 77 cats without mycosis, 72 (64.3%) and 35 (45.5%), respectively, were IFA reactive. No association was found between age, sex, FIV/FeLV and the presence of antibodies to Bartonella species. The results suggest that the study population can be considered a potential source of zoonotic infection for both diseases.
Assuntos
Anticorpos Antibacterianos/sangue , Infecções por Bartonella/veterinária , Doenças do Gato/microbiologia , Esporotricose/veterinária , Animais , Infecções por Bartonella/sangue , Infecções por Bartonella/imunologia , Brasil/epidemiologia , Doenças do Gato/epidemiologia , Gatos , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Vírus da Imunodeficiência Felina/imunologia , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/veterinária , Vírus da Leucemia Felina/imunologia , Masculino , Infecções por Retroviridae/epidemiologia , Infecções por Retroviridae/imunologia , Infecções por Retroviridae/veterinária , Esporotricose/epidemiologia , Esporotricose/microbiologia , Infecções Tumorais por Vírus/epidemiologia , Infecções Tumorais por Vírus/imunologia , Infecções Tumorais por Vírus/veterinária , ZoonosesRESUMO
Ovine lentivirus (OvLV) is a macrophage-tropic lentivirus found in many countries that causes interstitial pneumonia, mastitis, arthritis and cachexia in sheep. There is no preventive vaccine and no cure, but breed differences suggest marker-assisted selective breeding might improve odds of infection and control of OvLV post-infection. Although variants in TMEM154 have consistent association with odds of infection, no variant in any gene has been associated with host control of OvLV post-infection in multiple animal sets. Proviral concentration is a live-animal diagnostic measure of OvLV control post-infection related to severity of OvLV-induced lesions. A recent genome-wide association study identified a region including four zinc finger genes associated with proviral concentration in one Rambouillet flock. To refine this region, we tested additional variants and identified a small insertion/deletion variant near ZNF389 that showed consistent association with proviral concentration in three animal sets (P < 0.05). These animal sets contained Rambouillet, Polypay and crossbred sheep from multiple locations and management conditions. Strikingly, one flock had exceptionally high prevalence (>87%, including yearlings) and mean proviral concentration (>950 copies/µg), possibly due to needle sharing. The best estimate of proviral concentration by genotype, obtained from all 1310 OvLV-positive animals tested, showed insertion homozygotes had less than half the proviral concentration of other genotypes (P < 0.0001). Future work will test additional breeds, management conditions and viral subtypes, and identify functional properties of the haplotype this deletion variant tracks. To our knowledge, this is the first genetic variant consistently associated with host control of OvLV post-infection in multiple sheep flocks.
Assuntos
Resistência à Doença/genética , Infecções por Lentivirus/veterinária , Deleção de Sequência , Doenças dos Ovinos/genética , Animais , Genótipo , Infecções por Lentivirus/genética , Infecções por Lentivirus/imunologia , Lentivirus Ovinos-Caprinos/imunologia , Ovinos , Doenças dos Ovinos/imunologiaRESUMO
Small ruminant lentiviruses (SRLV) infect the monocyte/macrophage lineage inducing a long-lasting infection affecting body condition, production and welfare of sheep and goats all over the world. Macrophages play a pivotal role on the host's innate and adaptative immune responses against parasites by becoming differentially activated. Macrophage heterogeneity can tentatively be classified into classically differentiated macrophages (M1) through stimulation with IFN-γ displaying an inflammatory profile, or can be alternatively differentiated by stimulation with IL-4/IL-13 into M2 macrophages with homeostatic functions. Since infection by SRLV can modulate macrophage functions we explored here whether ovine and caprine macrophages can be segregated into M1 and M2 populations and whether this differential polarization represents differential susceptibility to SRLV infection. We found that like in human and mouse systems, ovine and caprine macrophages can be differentiated with particular stimuli into M1/M2 subpopulations displaying specific markers. In addition, small ruminant macrophages are plastic since M1 differentiated macrophages can express M2 markers when the stimulus changes from IFN-γ to IL-4. SRLV replication was restricted in M1 macrophages and increased in M2 differentiated macrophages respectively according to viral production. Identification of the infection pathways in macrophage populations may provide new targets for eliciting appropriate immune responses against SRLV infection.