RESUMO
The hypoxia-inducible factor 1 alpha (HIF1α) protein and the hypoxic microenvironment are critical for infection and pathogenesis by the oncogenic gammaherpesviruses (γHV), Kaposi sarcoma herpes virus (KSHV) and Epstein-Barr virus (EBV). However, understanding the role of HIF1α during the virus life cycle and its biological relevance in the context of host has been challenging due to the lack of animal models for human γHV. To study the role of HIF1α, we employed the murine gammaherpesvirus 68 (MHV68), a rodent pathogen that readily infects laboratory mice. We show that MHV68 infection induces HIF1α protein and HIF1α-responsive gene expression in permissive cells. siRNA silencing or drug-inhibition of HIF1α reduce virus production due to a global downregulation of viral gene expression. Most notable was the marked decrease in many viral genes bearing hypoxia-responsive elements (HREs) such as the viral G-Protein Coupled Receptor (vGPCR), which is known to activate HIF1α transcriptional activity during KSHV infection. We found that the promoter of MHV68 ORF74 is responsive to HIF1α and MHV-68 RTA. Moreover, Intranasal infection of HIF1αLoxP/LoxP mice with MHV68 expressing Cre- recombinase impaired virus expansion during early acute infection and affected lytic reactivation in the splenocytes explanted from mice. Low oxygen concentrations accelerated lytic reactivation and enhanced virus production in MHV68 infected splenocytes. Thus, we conclude that HIF1α plays a critical role in promoting virus replication and reactivation from latency by impacting viral gene expression. Our results highlight the importance of the mutual interactions of the oxygen-sensing machinery and gammaherpesviruses in viral replication and pathogenesis.
Assuntos
Regulação Viral da Expressão Gênica/fisiologia , Infecções por Herpesviridae/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Latência Viral/fisiologia , Replicação Viral/fisiologia , Animais , Camundongos , Rhadinovirus/metabolismoRESUMO
In the spring of 2017, a full-term infant with microcephaly was delivered in South Florida. During first trimester, the mother presented with fever, nausea, and vomiting. She reported no foreign travel for herself or her partner. The infant's neurologic, ophthalmologic, neuroradiologic, and audiologic findings were highly suggestive of congenital Zika syndrome (CZS), confirmed by IgM antibodies and plaque reduction neutralization test. New observations, including peripheral temporal retinal avascularity and peripapillary retinal nerve fiber layer thinning, are presented from this first known case of non-travel-associated CZS in the United States. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:e93-e98.].
Assuntos
Distrofias Hereditárias da Córnea/diagnóstico , Infecções Oculares Virais/diagnóstico , Microcefalia/diagnóstico , Doenças do Nervo Óptico/diagnóstico , Complicações Infecciosas na Gravidez , Infecção por Zika virus/diagnóstico , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Gravidez , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Ultrassonografia Pré-Natal , Estados Unidos , Adulto Jovem , Zika virus/genética , Infecção por Zika virus/congênitoRESUMO
The catalytically deficient ERBB3 strongly synergizes with the receptor tyrosine kinase ERBB2, and elevated levels represent an overall risk factor for unfavorable disease outcomes in breast cancer. Although itself not a target of pan-ERBB kinase inhibitors, it contributes to resistance in ERBB2-targeted treatment regiments. The steroidal lactone Withaferin A (WA) has established broad anticancer properties through several modes of action and was shown to be effective against triple-negative breast cancers at elevated concentrations. We found that ERBB2 overexpression does render cells hypersensitive to WA. Although ERBB2 downregulation is one aspect of WA treatment at high concentrations, it is not causal for the elevated sensitivity at lower dosages. Instead, WA targets the ability of ERBB3 to amplify ERBB2 signaling. ERBB3 receptor levels, constitutive phosphorylation of both ERBB3 and ERBB2, as well as signaling through AKT are eliminated by WA treatment. By targeting ERBB2/ERBB3 as a functional unit, it is also effective in cases in which ERBB2-directed inhibitors, such as lapatinib, alone show reduced potency. Hence, WA or derivatives thereof may present a low toxicity addition to ERBB2-targeting therapeutics, especially in cases in which ERBB3 involvement is driving resistance or reduced overall sensitivity. Mol Cancer Ther; 15(11); 2750-7. ©2016 AACR.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias da Mama/genética , Resistencia a Medicamentos Antineoplásicos/genética , Expressão Gênica , Receptor ErbB-2/genética , Receptor ErbB-3/genética , Vitanolídeos/farmacologia , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Receptor ErbB-2/metabolismo , Receptor ErbB-3/metabolismo , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/metabolismoRESUMO
Lytic replication of the Kaposi's sarcoma-associated herpesvirus (KSHV) is essential for the maintenance of both the infected state and characteristic angiogenic phenotype of Kaposi's sarcoma and thus represents a desirable therapeutic target. During the peak of herpesvirus lytic replication, viral glycoproteins are mass produced in the endoplasmic reticulum (ER). Normally, this leads to ER stress which, through an unfolded protein response (UPR), triggers phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF2α), resulting in inhibition of protein synthesis to maintain ER and cellular homeostasis. However, in order to replicate, herpesviruses have acquired the ability to prevent eIF2α phosphorylation. Here we show that clinically achievable nontoxic doses of the glucose analog 2-deoxy-d-glucose (2-DG) stimulate ER stress, thereby shutting down eIF2α and inhibiting KSHV and murine herpesvirus 68 replication and KSHV reactivation from latency. Viral cascade genes that are involved in reactivation, including the master transactivator (RTA) gene, glycoprotein B, K8.1, and angiogenesis-regulating genes are markedly decreased with 2-DG treatment. Overall, our data suggest that activation of UPR by 2-DG elicits an early antiviral response via eIF2α inactivation, which impairs protein synthesis required to drive viral replication and oncogenesis. Thus, induction of ER stress by 2-DG provides a new antiherpesviral strategy that may be applicable to other viruses.
Assuntos
Antivirais/farmacologia , Desoxiglucose/farmacologia , Retículo Endoplasmático/efeitos dos fármacos , Fator de Iniciação 2 em Eucariotos/antagonistas & inibidores , Herpesvirus Humano 8/efeitos dos fármacos , Resposta a Proteínas não Dobradas/efeitos dos fármacos , Proteínas Virais/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Retículo Endoplasmático/genética , Retículo Endoplasmático/virologia , Fator de Iniciação 2 em Eucariotos/genética , Fator de Iniciação 2 em Eucariotos/metabolismo , Expressão Gênica/efeitos dos fármacos , Herpesvirus Humano 8/fisiologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Humanos , Camundongos , Fosforilação , Ativação Transcricional/efeitos dos fármacos , Resposta a Proteínas não Dobradas/genética , Ensaio de Placa Viral , Proteínas Virais/biossíntese , Proteínas Virais/genética , Ativação Viral/efeitos dos fármacos , Latência Viral/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
BACKGROUND: Defects in the immune system may affect vaccine responsiveness. Because of the splenic hypofunction and abnormal opsonic activity, it was unknown whether patients with sickle cell disease (SCD) would respond appropriately to H1N1 vaccination. The objective of this study was to assess seroprotective post-vaccine H1N1 antibody response in children with SCD. PROCEDURE: Serum antibody titers were measured by hemagglutination inhibition and microneutralization (MN) assays. Correlations were established between clinical and treatment parameters and immune response. RESULTS: Twenty-nine of 38 (76.3%) subjects (mean age 11 ± 5.4 years) had durable protective antibody titers 8 ± 1.6 months (range 5-12 months) post-vaccination. Lessened immune response was not associated with time interval from vaccination, splenectomy, or hydroxyurea treatment. Lack of antibody response was associated with age less than 3 years and treatment with chronic transfusions. Of the nine non-responders, seven were on chronic transfusions (39% unresponsiveness rate in the transfused group). The difference in the number of patients with seropositivity between the non-transfused and the transfused groups was statistically significant (P = 0.039). CONCLUSIONS: Most subjects were able to mount an influenza-specific antibody response against the inactivated H1N1 vaccine. Similar to the general population, children less than 3 years were less likely to respond. In addition, patients on chronic transfusions were less likely to respond when compared to non-transfused children. Clinicians should be aware of the possibility of decreased vaccine response in patients with SCD on chronic transfusions. We postulate that transfusion-related immunomodulation (TRIM) may be related to decreased response.
Assuntos
Anemia Falciforme/imunologia , Anticorpos Antivirais/sangue , Transfusão de Sangue , Vírus da Influenza A Subtipo H1N1/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Adolescente , Anemia Falciforme/sangue , Anemia Falciforme/terapia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Esplenectomia , Vacinas de Produtos Inativados , Adulto JovemRESUMO
We have documented previously that a multiple chaperone protein vaccine termed chaperone-rich cell lysate (CRCL) promotes tumor-specific T-cell responses leading to cancer regression in several mouse tumor models. We report here that CRCL vaccine generated from a mouse breast cancer (TUBO, HER2/neu positive) is also capable of eliciting humoral immunity. Administration of TUBO CRCL triggered anti-HER2/neu antibody production and delayed the progression of established tumors. This antitumor activity can be transferred through the serum isolated from TUBO CRCL-immunized animals and involved both B cells and CD4(+) T lymphocytes. Further evaluation of the mechanisms underlying TUBO CRCL-mediated humoral immunity highlighted the role of antibody-dependent cell-mediated cytotoxicity. These results suggest that tumor-derived CRCL vaccine has a wider applicability as a cancer vaccine because it can target both T-cell- and B-cell-specific responses and may represent a promising approach for the immunotherapy of cancer.
Assuntos
Anticorpos Antineoplásicos/biossíntese , Vacinas Anticâncer/imunologia , Modelos Animais de Doenças , Neoplasias Mamárias Experimentais/imunologia , Chaperonas Moleculares/metabolismo , Animais , Citotoxicidade Celular Dependente de Anticorpos , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Soros Imunes , Neoplasias Mamárias Experimentais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos TransgênicosRESUMO
CD4(+)CD25(+) regulatory T lymphocytes (Tregs) critically contribute to the mechanisms of cancer-induced tolerance. These cells suppress anti-tumoral CD8(+) and CD4(+) T lymphocytes and can also restrain the function of APCs. We have previously documented the immunostimulatory effects of a chaperone-rich cell lysate (CRCL) anti-cancer vaccine. Tumor-derived CRCL induces tumor immunity in vivo, partly by promoting dendritic cell (DC) and macrophage activation. In the current study, we evaluated the effects of CD4(+)CD25(+)forkhead box P3(+) Tregs isolated from mice bearing 12B1 bcr-abl(+) leukemia on DC and macrophages that had been activated by 12B1-derived CRCL. CRCL-activated DC and macrophages resisted Treg suppression, as the production of proinflammatory cytokines, the activation of transcription factor NF-kappaB, and their immunostimulatory potential was unaffected by Tregs. Our results thus highlight CRCL as a powerful adjuvant endowed with the capacity to overcome tumor-induced Treg-inhibitory effects on APCs.
Assuntos
Células Dendríticas/imunologia , Linfócitos T Reguladores/imunologia , Animais , Células da Medula Óssea/imunologia , Linhagem Celular Tumoral , Terapia de Imunossupressão , Leucemia Experimental/patologia , Ativação Linfocitária , Depleção Linfocítica , Macrófagos Peritoneais/imunologia , Camundongos , NF-kappa B/metabolismoRESUMO
CD4(+)CD25(+) regulatory T cells have been characterized as a critical population of immunosuppressive cells. They play a crucial role in cancer progression by inhibiting the effector function of CD4(+) or CD8(+) T lymphocytes. However, whether regulatory T lymphocytes that expand during tumor progression can modulate dendritic cell function is unclear. To address this issue, we have evaluated the inhibitory potential of CD4(+)CD25(+) regulatory T cells from mice bearing a BCR-ABL(+) leukemia on bone marrow-derived dendritic cells. We present data demonstrating that CD4(+)CD25(+)FoxP3(+) regulatory T cells from tumor-bearing animals impede dendritic cell function by down-regulating the activation of the transcription factor NF-kappaB. The expression of the co-stimulatory molecules CD80, CD86 and CD40, the production of TNF-alpha, IL-12, and CCL5/RANTES by the suppressed DC is strongly down-regulated. The suppression mechanism requires TGF-beta and IL-10 and is associated with induction of the Smad signaling pathway and activation of the STAT3 transcription factor.
Assuntos
Antígenos CD4/imunologia , Células Dendríticas/fisiologia , Terapia de Imunossupressão , Interleucina-10/metabolismo , Subunidade alfa de Receptor de Interleucina-2/imunologia , Linfócitos T Reguladores/imunologia , Fator de Crescimento Transformador beta/metabolismo , Animais , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Células da Medula Óssea , Feminino , Citometria de Fluxo , Fatores de Transcrição Forkhead/metabolismo , Proteínas de Fusão bcr-abl/metabolismo , Leucemia/imunologia , Leucemia/patologia , Leucemia/prevenção & controle , Ativação Linfocitária , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/metabolismo , Linfócitos do Interstício Tumoral/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , NF-kappa B/metabolismo , Fosforilação , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de SinaisRESUMO
Tumor derived chaperone-rich cell lysate (CRCL) when isolated from tumor tissues is a potent vaccine that contains at least 4 of the highly immunogenic heat shock proteins (HSP) such as HSP70, HSP90, glucose related protein 94 and calreticulin. We have previously documented that CRCL provides both a source of tumor antigens and danger signals triggering dendritic cell (DC) activation. Immunization with tumor derived CRCL elicits tumor-specific T cell responses leading to tumor regression. In the current study, we further dissect the mechanisms by which CRCL simulates the immune system, and demonstrate that natural killer (NK) cells are required for effective antitumor effects to take place. Our results illustrate that CRCL directly stimulates proinflammatory cytokine and chemokine production by NK cells, which may lead to activation and recruitment of macrophages at the tumor site. Thus, this report provides further insight into the function of CRCL as an immunostimulant against cancer.
Assuntos
Vacinas Anticâncer/administração & dosagem , Células Matadoras Naturais/imunologia , Neoplasias Experimentais/imunologia , Animais , Vacinas Anticâncer/imunologia , Citocinas/biossíntese , Citocinas/fisiologia , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Ativação de Macrófagos , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
OBJECTIVE: To quantify the immune response of WASP- mice to three different pathogens: influenza A virus, Streptococcus pneumoniae, and Mycobacterium bovis. METHODS: Primary and secondary T-cell responses to influenza A virus were quantified via tetramer assays. Viral clearance from lung was also measured. Lethality of intranasal inoculation with luminescent S. pneumoniae was assessed by dose escalation and direct luminescence imaging. After intravenous inoculation with M. bovis, residual mycobacteria in lung, liver, and spleen were measured by standard culture methods. RESULTS: The reduced secondary T-cell response to influenza A virus correlates with a relative but not absolute loss of splenic T and B cells similar to that seen in clinical Wiskott-Aldrich Syndrome (WAS), and slower clearance of virus from lung. The reduced magnitude of the secondary T-cell response correlates with a progressive loss of influenza-specific T cells after primary inoculation. WASP- mice show an increased susceptibility to lethal pneumonia after intranasal inoculation with S. pneumoniae, which is among the most frequent causes of clinical complications in WAS patients. WASP- mice clear M. bovis bacille Calmette-Guerin (BCG) more slowly from lung, liver, and spleen. Bone marrow-derived macrophages, however, show normal ex vivo cytokine secretion in response to M. bovis. CONCLUSIONS: These results demonstrate that WASP- mice are functionally immunodeficient in regard to three different pathogens, and provide relevant end points for the study of treatment modalities in this model. They also suggest a specific physiologic mechanism, failure to accumulate memory T cells, for at least one of the defective immune responses.
Assuntos
Imunidade , Proteínas/imunologia , Linfócitos T/imunologia , Animais , Citocinas/metabolismo , Suscetibilidade a Doenças , Vírus da Influenza A/imunologia , Fígado/microbiologia , Pulmão/microbiologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Mycobacterium bovis/imunologia , Proteínas/genética , Baço/microbiologia , Streptococcus pneumoniae/imunologia , Proteína da Síndrome de Wiskott-AldrichRESUMO
Can CD4(+) and CD8(+) "memory" T cells that are generated and maintained in the context of low-level virus persistence protect, in the absence of antibody, against a repeat challenge with the same pathogen? Although immune T cells exert effective, long-term control of a persistent gamma-herpesvirus (gammaHV68) in Ig(-/-) microMT mice, subsequent exposure to a high dose of the same virus leads to further low-level replication in the lung. This lytic phase in the respiratory tract is dealt with effectively by the recall of memory T cells induced by a gammaHV68 recombinant (M3LacZ) that does not express the viral M3 chemokine binding protein. At least for the CD8(+) response, greater numbers of memory T cells confer enhanced protection in the M3LacZ-immune mice. However, neither WT gammaHV68 nor the minimally persistent M3LacZ primes the T cell response to the extent that a WT gammaHV68 challenge fails to establish latency in the microMT mice. Memory CD4(+) and CD8(+) T cells thus act together to limit gammaHV68 infection but are unable to provide absolute protection against a high-dose, homologous challenge.
Assuntos
Gammaherpesvirinae/imunologia , Imunoglobulinas/deficiência , Memória Imunológica/imunologia , Superinfecção/imunologia , Superinfecção/virologia , Linfócitos T/imunologia , Linfócitos T/virologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/virologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/virologia , Imunização Secundária , Imunoglobulinas/genética , Pulmão/virologia , Camundongos , Baço/imunologia , Baço/virologia , Latência Viral/imunologiaRESUMO
We have investigated the interactions and consequences of superinfecting and coreplication of human papillomavirus (HPV) and herpes simplex virus (HSV) in human epithelial organotypic (raft) culture tissues. In HPV-positive tissues, HSV infection and replication induced significant cytopathic effects (CPE), but the tissues were able to recover and maintain a certain degree of tissue integrity and architecture. HPV31b not only maintained the episomal state of its genomic DNA but also maintained its genomic copy number even during times of extensive HSV-induced CPE. E2 transcripts encoded by HPV31b were undetectable even though HPV31b replication was maintained in HSV- infected raft tissues. Expression of HPV31b oncogenes (E6 and E7) was also repressed but to a lesser degree than was E2 expression. The extent of CPE induced by HSV is dependent on the magnitude of HPV replication and gene expression at the time of HSV infection. During active HSV infection, HPV maintains its genomic copy number even though genes required for its replication were repressed. These studies provide new insight into the complex interaction between two common human sexually transmitted viruses in an in vitro system, modeling their natural host tissue in vivo.
Assuntos
Herpesvirus Humano 1/patogenicidade , Herpesvirus Humano 2/patogenicidade , Papillomaviridae/patogenicidade , Replicação Viral , Diferenciação Celular , Linhagem Celular Transformada , Células Cultivadas , Técnicas de Cultura , Efeito Citopatogênico Viral , DNA Viral/análise , Células Epiteliais , Feminino , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/fisiologia , Herpesvirus Humano 2/genética , Herpesvirus Humano 2/fisiologia , Humanos , Imuno-Histoquímica , Papillomaviridae/genética , Papillomaviridae/fisiologia , Células Tumorais Cultivadas , Proteínas Virais/metabolismoRESUMO
The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder characterized by immune dysfunction, thrombocytopenia, and eczema. We used a murine model created by knockout of the WAS protein gene (WASP) to evaluate the potential of gene therapy for WAS. Lethally irradiated, male WASP- animals that received transplants of mixtures of wild type (WT) and WASP- bone marrow cells demonstrated enrichment of WT cells in the lymphoid and myeloid lineages with a progressive increase in the proportion of WT T-lymphoid and B-lymphoid cells. WASP- mice had a defective secondary T-cell response to influenza virus which was normalized in animals that received transplants of 35% or more WT cells. The WASP gene was inserted into WASP- bone marrow cells with a bicistronic oncoretroviral vector also encoding green fluorescent protein (GFP), followed by transplantation into irradiated male WASP- recipients. There was a selective advantage for gene-corrected cells in multiple lineages. Animals with higher proportions of GFP+ T cells showed normalization of their lymphocyte counts. Gene-corrected, blood T cells exhibited full and partial correction, respectively, of their defective proliferative and cytokine secretory responses to in vitro T-cell-receptor stimulation. The defective secondary T-cell response to influenza virus was also improved in gene-corrected animals.
Assuntos
Terapia Genética/métodos , Orthomyxoviridae/imunologia , Proteínas/administração & dosagem , Linfócitos T/imunologia , Síndrome de Wiskott-Aldrich/terapia , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Transplante de Medula Óssea , Vetores Genéticos , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Imunidade Celular , Masculino , Camundongos , Camundongos Knockout , Infecções por Orthomyxoviridae/complicações , Infecções por Orthomyxoviridae/imunologia , Proteínas/genética , Retroviridae/genética , Transdução Genética/métodos , Resultado do Tratamento , Síndrome de Wiskott-Aldrich/complicações , Síndrome de Wiskott-Aldrich/imunologia , Proteína da Síndrome de Wiskott-AldrichRESUMO
The consequences for the long-term maintenance of virus-specific CD8+-T-cell memory have been analyzed experimentally for sequential respiratory infections with readily eliminated (influenza virus) and persistent (gammaherpesvirus 68 [gammaHV68]) pathogens. Sampling a broad range of tissue sites established that the numbers of CD8+ T cells specific for the prominent influenza virus D(b)NP(366) epitope were reduced by about half in mice that had been challenged 100 days previously with gammaHV68, though the prior presence of a large CD8+ D(b)NP366+ population caused no selective defect in the gammaHV68-specific CD8+ K(b)p79+ response. Conversely, mice that had been primed and boosted to generate substantial gammaHV68-specific CD8+ D(b)p56+ populations did not show any decrease in prevalence for this set of CD8+ memory cytotoxic T lymphocytes (CTL) at 200 days after respiratory exposure to an influenza A virus. However, in both experiments, the total magnitude of the CD8+-T-cell pool was significantly diminished in those that had been infected with gammaHV68 and the influenza A virus. The broader implications of these findings, especially under conditions of repeated exposure to unrelated pathogens, are explored with a mathematical model which emphasizes that the immune effector and memory "phenome" is a function of the overall infection experience of the individual.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Gammaherpesvirinae/patogenicidade , Memória Imunológica , Vírus da Influenza A/imunologia , Infecções por Orthomyxoviridae/imunologia , Animais , Feminino , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Modelos Imunológicos , Infecções por Orthomyxoviridae/virologia , Fatores de TempoRESUMO
The murine gammaherpesvirus-68 kills I-A(b-/-) mice despite the presence of virus-specific CD8+ cytotoxic T lymphocytes (CTL). This has raised the possibility that these CTL are functionally abnormal. Here, no difference was observed between I-A(b-/-) mice and I-A(b+/+) controls in virus-specific CTL function, T cell receptor usage, or surface phenotype. Thus CTL immunity was independent of CD4+ T cells in a chronic herpesvirus infection, but was still inadequate to control virus replication.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Gammaherpesvirinae/patogenicidade , Infecções por Herpesviridae/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Antígenos H-2/metabolismo , Infecções por Herpesviridae/virologia , Antígeno de Histocompatibilidade H-2D , Camundongos , Fragmentos de Peptídeos/metabolismo , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismoRESUMO
Mice (I-A(b-/-)) that lack CD4(+) T cells remain healthy for at least three months after respiratory exposure to the murine gamma-herpesvirus 68 (gammaHV68), then succumb with symptoms of chronic wasting disease. Postexposure challenge of gammaHV68-infected I-A(b+/+) and I-A(b-/-) mice with a recombinant vaccinia virus (Vacc-p56) expressing an antigenic gammaHV68 peptide caused a massive increase in the numbers of D(b)p56-specific CD8(+) T cells. Previous experiments showed that, despite the large numbers of potential CTL effectors, there was little effect on the long-term survival of the CD4-deficient group and no diminution in the level of persistent virus shedding and latency. Comparison of the expanded CD8(+)D(b)p56(+) sets in the I-A(b+/+) and I-A(b-/-) mice indicated that these two T cell populations were not identical. More CD69(high)CD8(+) D(b)p56(+) T cells were found in the CD4-deficient mice, an effect that might be thought to reflect higher Ag load. By contrast, the mean fluorescence intensity of staining for the CD44 glycoprotein was diminished on CD8(+)D(b)p56(+) T cells from the I-A(b-/-) group, the level of CTL activity was lower on a per cell basis, and the relative prevalence of IFN-gamma(+)TNF-alpha(+) T cells detected after in vitro stimulation with the p56 peptide was decreased. Given that this experimental system provides an accessible model for evaluating postexposure vaccination protocols that might be used in diseases like HIV/AIDS, the further need is to clarify the underlying molecular mechanisms and the relative significance of lack of CD4(+) T help vs higher Ag load for these expanded CD8(+) effector populations.