RESUMO
PURPOSE: This pilot study aimed on generating insight on alterations in circulating immune cells during the use of FOLFIRINOX and gemcitabine/nab-paclitaxel in pancreatic ductal adenocarcinoma (PDAC). PATIENTS AND METHODS: Peripheral blood mononuclear cells were isolated before and 30 days after initiation of chemotherapy from 20 patients with advanced PDAC. Regulatory T cells (FoxP3+) and immune checkpoints (PD-1 and TIM-3) were analyzed by flow cytometry and immunological changes were correlated with clinical outcome. RESULTS: Heterogeneous changes during chemotherapy were observed in circulating T-cell subpopulations with a pronounced effect on PD-1+ CD4+/CD8+ T cells. An increase in FoxP3+ or PD-1+ T cells had no significant effect on survival. An increase in TIM3+/CD8+ (but not TIM3+/CD4+) T cells was associated with a significant inferior outcome: median progression-free survival in the subgroup with an increase of TIM-3+/CD8+ T cells was 6.0 compared to 14.0 months in patients with a decrease/no change (p = 0.026); corresponding median overall survival was 13.0 and 20.0 months (p = 0.011), respectively. CONCLUSIONS: Chemotherapy with FOLFIRNOX or gemcitabine/nab-paclitaxel induces variable changes in circulating T-cell populations that may provide prognostic information in PDAC.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma Ductal Pancreático/tratamento farmacológico , Proteínas de Checkpoint Imunológico/efeitos dos fármacos , Neoplasias Pancreáticas/tratamento farmacológico , Linfócitos T Reguladores/efeitos dos fármacos , Idoso , Albuminas/uso terapêutico , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/química , Linfócitos T CD8-Positivos/efeitos dos fármacos , Carcinoma Ductal Pancreático/imunologia , Desoxicitidina/análogos & derivados , Desoxicitidina/uso terapêutico , Feminino , Fluoruracila/uso terapêutico , Fatores de Transcrição Forkhead , Receptor Celular 2 do Vírus da Hepatite A/análise , Humanos , Proteínas de Checkpoint Imunológico/análise , Irinotecano/uso terapêutico , Leucovorina/uso terapêutico , Masculino , Pessoa de Meia-Idade , Oxaliplatina/uso terapêutico , Paclitaxel/uso terapêutico , Neoplasias Pancreáticas/imunologia , Projetos Piloto , Receptor de Morte Celular Programada 1/análise , Receptor de Morte Celular Programada 1/efeitos dos fármacos , Intervalo Livre de Progressão , Estudos Prospectivos , Linfócitos T Reguladores/química , GencitabinaRESUMO
OBJECTIVE: To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. METHODS: Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. RESULTS: Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. CONCLUSION: Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.
Assuntos
Líquido Ascítico/patologia , Endometriose/patologia , Endométrio/patologia , Linfócitos T Reguladores/química , Receptores Toll-Like/análise , Adolescente , Adulto , Líquido Ascítico/imunologia , Índice de Massa Corporal , Estudos de Casos e Controles , Endometriose/imunologia , Endométrio/imunologia , Feminino , Humanos , Valores de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estatísticas não Paramétricas , Linfócitos T Reguladores/imunologia , Escala Visual Analógica , Adulto JovemRESUMO
ABSTRACT Objective To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. Methods Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. Results Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. Conclusion Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.
RESUMO Objetivo Analisar e comparar a expressão de receptores do tipo Toll por células T reguladoras presentes no líquido peritoneal de pacientes com endometriose. Métodos Células T reguladoras foram isoladas do líquido peritoneal de mulheres com e sem endometriose, coletadas durante a cirurgia, e o RNAm foi extraído para análise da expressão de receptores do tipo Toll por reação em cadeia da polimerase com transcriptase reversa. Resultados Pacientes com endometriose apresentaram células T reguladoras expressando maior número e variedade de Toll por células quando comparadas com T reguladoras de pacientes do Grupo Controle. Receptores do tipo Toll-1 e receptores do tipo Toll-2 foram expressos em ambos os grupos. Todos os outros tipos de receptores Toll foram encontrados expressos apenas em células T reguladoras do grupo com endometriose. Conclusão Pacientes com endometriose apresentaram células T reguladoras peritoneais expressando vários tipos de receptores tipo Toll.
Assuntos
Humanos , Feminino , Adolescente , Adulto , Adulto Jovem , Líquido Ascítico/patologia , Linfócitos T Reguladores/química , Endometriose/patologia , Endométrio/patologia , Receptores Toll-Like/análise , Valores de Referência , Líquido Ascítico/imunologia , Índice de Massa Corporal , Estudos de Casos e Controles , Linfócitos T Reguladores/imunologia , Estatísticas não Paramétricas , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Endometriose/imunologia , Endométrio/imunologia , Escala Visual AnalógicaRESUMO
Human Leukocyte Antigen G (HLA-G) is a molecule involved in the tumor immunosuppression and also in the generation of regulatory T (Treg) cells, thus leading to evasion to the immune system host, and consequently, contributing to tumor progression in several cancers. The aim of this study was to evaluate the immunoexpression of HLA-G by tumor cells and FoxP3+ Treg cells in 25 oral tongue squamous cell carcinomas (SCCs) and 25 lower lip SCCs and analyze their relationship with clinical parameters. HLA-G expression was higher in oral tongue SCCs than in lower lip SCCs. In oral tongue SCCs and lower lip SCCs, no association between HLA-G expression and clinical parameters (tumor size, lymph node status, distant metastasis, and clinical stage) was verified (P>0.05). FoxP3+ Treg cells were detected along the tumor invasive front in all cases of oral tongue and lower lip SCCs. In oral tongue SCC cases, the number of Treg cells tended to be higher in smaller tumors, tumors without regional lymph node metastasis, and tumors in early clinical stages, but the difference was not statistically significant (P>0.05). A significant positive correlation was found between the expression of HLA-G by neoplastic cells and Treg cells in lower lip SCCs (p = 0.008). Our findings suggest the involvement of HLA-G and Treg cells in the modulation of immune responses in oral tongue and lower lip SCCs. This interaction between HLA-G and Treg cells may represent an evasion mechanism in these malignancies.
Assuntos
Carcinoma de Células Escamosas/patologia , Fatores de Transcrição Forkhead/análise , Antígenos HLA-G/análise , Neoplasias Labiais/patologia , Linfócitos T Reguladores/química , Neoplasias da Língua/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Valores de Referência , Estatísticas não Paramétricas , Linfócitos T Reguladores/patologia , Carga TumoralRESUMO
Infection with human papillomavirus (HPV) is the main cause of cervical cancer. Viral persistence is considered the main risk factor for neoplastic progression and evidence suggests that regulatory T cells (Treg) play an important role in the failure of viral elimination. The aim of this study was to detect phenotypic markers of Treg and cytokines interleukin (IL)-10 and transforming growth factor (TGF)-ß, in the cervical microenvironment of HPV-infected patients. One hundred and one samples of uterine cervix embedded in paraffin were analyzed. We used immunohistochemistry to examine the coexpression of the CD25/FOXP3 and CD4/TGF-ß markers, and the expression of GITR and IL-10 in cells present in the cervical stroma. We detected a microenvironment composed of high proportions of CD25+ FOXP3+ , CD4+ TGFß+ , IL-10+ , and GITR+ cells in samples with high viral loads and severe lesions of HPV-infected patients. The abundance of these markers, indicative of the presence of Treg cells and immunosuppressive cytokines, was significantly associated with severe lesions and elevated viral loads in the examined samples. These results suggest that Treg cells may be involved in maintaining a microenvironment favorable for viral persistence and neoplastic progression. Our findings support those of previous studies that suggested that these markers could be used to predict HPV persistence and neoplastic progression, and as potential targets for immune response modulation.
Assuntos
Colo do Útero/patologia , Citocinas/análise , Infecções por Papillomavirus/patologia , Subpopulações de Linfócitos T/química , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/química , Linfócitos T Reguladores/imunologia , Adolescente , Adulto , Idoso , Colo do Útero/virologia , Feminino , Humanos , Pessoa de Meia-Idade , Papillomaviridae/imunologia , Infecções por Papillomavirus/virologia , Carga Viral , Adulto JovemRESUMO
Abstract: Human Leukocyte Antigen G (HLA-G) is a molecule involved in the tumor immunosuppression and also in the generation of regulatory T (Treg) cells, thus leading to evasion to the immune system host, and consequently, contributing to tumor progression in several cancers. The aim of this study was to evaluate the immunoexpression of HLA-G by tumor cells and FoxP3+ Treg cells in 25 oral tongue squamous cell carcinomas (SCCs) and 25 lower lip SCCs and analyze their relationship with clinical parameters. HLA-G expression was higher in oral tongue SCCs than in lower lip SCCs. In oral tongue SCCs and lower lip SCCs, no association between HLA-G expression and clinical parameters (tumor size, lymph node status, distant metastasis, and clinical stage) was verified (P>0.05). FoxP3+ Treg cells were detected along the tumor invasive front in all cases of oral tongue and lower lip SCCs. In oral tongue SCC cases, the number of Treg cells tended to be higher in smaller tumors, tumors without regional lymph node metastasis, and tumors in early clinical stages, but the difference was not statistically significant (P>0.05). A significant positive correlation was found between the expression of HLA-G by neoplastic cells and Treg cells in lower lip SCCs (p = 0.008). Our findings suggest the involvement of HLA-G and Treg cells in the modulation of immune responses in oral tongue and lower lip SCCs. This interaction between HLA-G and Treg cells may represent an evasion mechanism in these malignancies.
Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Neoplasias Labiais/patologia , Neoplasias da Língua/patologia , Carcinoma de Células Escamosas/patologia , Linfócitos T Reguladores/química , Fatores de Transcrição Forkhead/análise , Antígenos HLA-G/análise , Valores de Referência , Imuno-Histoquímica , Linfócitos T Reguladores/patologia , Estatísticas não Paramétricas , Carga Tumoral , Pessoa de Meia-Idade , Estadiamento de NeoplasiasRESUMO
The aim of this study was to determine the number of FoxP3+ regulatory T (Treg) cells in the microenvironment of lower lip squamous cell carcinomas (LLSCCs) and to correlate the findings with clinicopathological parameters (tumor size/extent, regional lymph node metastasis, clinical stage, and histopathological grade of malignancy). Fifty cases of LLSCC were selected. Lymphocytes exhibiting nuclear immunostaining for FoxP3 were quantified in 10 microscopic fields at the deep invasive front of LLSCCs. The results were analyzed statistically using the nonparametric Mann-Whitney test and Fisher's exact test. FoxP3+ lymphocytes were observed in all cases studied. The number of these cells tended to be higher in smaller tumors, tumors without regional lymph node metastasis, and tumors in early clinical stages, but the difference was not statistically significant (p > 0.05). Low-grade tumors contained a larger number of FoxP3+ lymphocytes than high-grade tumors (p = 0.019). Tumors with an intense inflammatory infiltrate exhibited a larger number of Treg cells (p = 0.035). On the other hand, the number of FoxP3+ lymphocytes was smaller in tumors arranged in small cell clusters (p = 0.003). No significant differences in the number of FoxP3+ lymphocytes were observed according to the degree of keratinization (p = 0.525) or nuclear pleomorphism (p = 0.343). The results suggest the participation of Treg cells in immune and inflammatory responses in the microenvironment of LLSCCs. These cells may play a more important role in early stages rather than in advanced stages of lip carcinogenesis.
Assuntos
Carcinoma de Células Escamosas/patologia , Fatores de Transcrição Forkhead/análise , Neoplasias Labiais/patologia , Linfócitos T Reguladores/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Contagem de Células , Feminino , Humanos , Imuno-Histoquímica , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Valores de Referência , Linfócitos T Reguladores/patologia , Carga Tumoral , Microambiente TumoralRESUMO
Several CD4+ T cell subtypes contribute to immune homeostasis in malaria, but the markers that define the main suppressive T cell subsets induced by this infection remain largely unknown. Here we provide a detailed phenotypic characterization of immunoregulatory CD4+ T cell populations in uncomplicated human malaria. We found an increased proportion of CD4+ T cells expressing CTLA-4, OX40, GITR, TNFRII, and CD69 in acute-phase single-species infections with Plasmodium vivax, Plasmodium falciparum, or both. Such an increase was not proportional to parasite density in P. vivax infections, and did not persist after parasite clearance. Significantly, less than 10% of CD4+ T cells expressing these regulatory molecules had the classical T regulatory (Treg) phenotype (CD4+CD25+CD127-FoxP3+). Two major Treg cell subpopulations, which together accounted for 19-23% of all Treg cells circulating in malaria patients, expressed surface receptors with opposing regulatory functions, either CTLA-4 or OX40. OX40+ Treg cells outnumbered their CTLA-4+ counterparts (1.8:1) during acute P. vivax infection, while a more balanced ratio (1.3:1) was observed following parasite clearance These data reveal new players in the complex CD4+ Treg cell network that maintains immune homeostasis in malaria and suggest potential targets for therapeutic interventions to improve parasite-specific effector immune responses.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Antígeno CTLA-4/análise , Malária Falciparum/patologia , Malária Vivax/patologia , Receptores OX40/análise , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Linfócitos T CD4-Positivos/química , Feminino , Homeostase , Humanos , Malária Falciparum/imunologia , Malária Vivax/imunologia , Masculino , Pessoa de Meia-Idade , Subpopulações de Linfócitos T/química , Linfócitos T Reguladores/química , Adulto JovemRESUMO
Abstract: The aim of this study was to determine the number of FoxP3+ regulatory T (Treg) cells in the microenvironment of lower lip squamous cell carcinomas (LLSCCs) and to correlate the findings with clinicopathological parameters (tumor size/extent, regional lymph node metastasis, clinical stage, and histopathological grade of malignancy). Fifty cases of LLSCC were selected. Lymphocytes exhibiting nuclear immunostaining for FoxP3 were quantified in 10 microscopic fields at the deep invasive front of LLSCCs. The results were analyzed statistically using the nonparametric Mann-Whitney test and Fisher's exact test. FoxP3+ lymphocytes were observed in all cases studied. The number of these cells tended to be higher in smaller tumors, tumors without regional lymph node metastasis, and tumors in early clinical stages, but the difference was not statistically significant (p > 0.05). Low-grade tumors contained a larger number of FoxP3+ lymphocytes than high-grade tumors (p = 0.019). Tumors with an intense inflammatory infiltrate exhibited a larger number of Treg cells (p = 0.035). On the other hand, the number of FoxP3+ lymphocytes was smaller in tumors arranged in small cell clusters (p = 0.003). No significant differences in the number of FoxP3+ lymphocytes were observed according to the degree of keratinization (p = 0.525) or nuclear pleomorphism (p = 0.343). The results suggest the participation of Treg cells in immune and inflammatory responses in the microenvironment of LLSCCs. These cells may play a more important role in early stages rather than in advanced stages of lip carcinogenesis.
Assuntos
Humanos , Masculino , Feminino , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/patologia , Linfócitos T Reguladores/química , Fatores de Transcrição Forkhead/análise , Valores de Referência , Neoplasias Labiais/patologia , Imuno-Histoquímica , Contagem de Células , Linfócitos T Reguladores/patologia , Carga Tumoral , Microambiente Tumoral , Gradação de Tumores , Metástase Linfática , Pessoa de Meia-Idade , Estadiamento de NeoplasiasRESUMO
Paracoccidioidomycosis (PCM), is a pulmonary fungal disease whose severity depends on the adequate development of T cell immunity. Although regulatory T (Treg) cells were shown to control immunity against PCM, deleterious or protective effects were described in different experimental settings. To clarify the function of Treg cells in pulmonary PCM, loss-and gain-of-function approaches were performed with Foxp3GFP knock-in mice and immunodeficient Rag1-/- mice, respectively, which were intratracheally infected with 106 yeast cells. The activity of Foxp3-expressing Treg cells in pulmonary PCM was determined in Foxp3GFP transgenic mice. First, it was verified that natural Treg cells migrate to the lungs of infected mice, where they become activated. Depletion of Treg cells led to reduced fungal load, diminished pathogen dissemination and increased Th1/Th2/Th17 immunity. Further, adoptive transfer of diverse T cell subsets to Rag1-/- mice subsequently infected by the pulmonary route demonstrated that isolated CD4+Foxp3+ Treg cells were able to confer some degree of immunoprotection and that CD4+Foxp3- T cells alone reduced fungal growth and enhanced T cell immunity, but induced vigorous inflammatory reactions in the lungs. Nevertheless, transfer of Treg cells combined with CD4+Foxp3- T cells generated more efficient and balanced immune Th1/Th2/Th17 responses able to limit pathogen growth and excessive tissue inflammation, leading to regressive disease and increased survival rates. Altogether, these loss- and gain-of-function approaches allow us to clearly demonstrate the dual role of Treg cells in pulmonary PCM, their deleterious effects by impairing T cell immunity and pathogen eradication, and their protective role by suppressing exacerbated tissue inflammation.