RESUMO
Treatment with anti-CD20, used in many diseases in which B cells play a pathogenic role, has been associated with susceptibility to intracellular infections. Here, we studied the effect of anti-CD20 injection on CD8+ T cell immunity using an experimental model of Trypanosoma cruzi infection, in which CD8+ T cells play a pivotal role. C57BL/6 mice were treated with anti-CD20 for B cell depletion prior to T. cruzi infection. Infected anti-CD20-treated mice exhibited a CD8+ T cell response with a conserved expansion phase followed by an early contraction, resulting in a strong reduction in total and parasite-specific CD8+ T cell numbers at 20 days postinfection. Anti-CD20 injection increased the frequency of apoptotic CD8+ T cells, decreased the number of effector and memory CD8+ T cells, and reduced the frequency of proliferating and cytokine-producing CD8+ T cells. Accordingly, infected anti-CD20-treated mice presented lower cytotoxicity of T. cruzi peptide-pulsed target cells in vivo All of these alterations in CD8+ T cell immunity were associated with increased tissue parasitism. Anti-CD20 injection also dampened the CD8+ T cell response, when this had already been generated, indicating that B cells were involved in the maintenance rather than the induction of CD8+ T cell immunity. Anti-CD20 injection also resulted in a marked reduction in the frequency of interleukin-6 (IL-6)- and IL-17A-producing cells, and recombinant IL-17A (rIL-17A) injection partially restored the CD8+ T cell response in infected anti-CD20-treated mice. Thus, anti-CD20 reduced CD8+ T cell immunity, and IL-17A is a candidate for rescuing deficient responses either directly or indirectly.IMPORTANCE Monoclonal antibody targeting the CD20 antigen on B cells is used to treat the majority of non-Hodgkin lymphoma patients and some autoimmune disorders. This therapy generates adverse effects, notably opportunistic infections and activation of viruses from latency. Here, using the infection murine model with the intracellular parasite Trypanosoma cruzi, we report that anti-CD20 treatment affects not only B cell responses but also CD8+ T cell responses, representing the most important immune effectors involved in control of intracellular pathogens. Anti-CD20 treatment, directly or indirectly, affects cytotoxic T cell number and function, and this deficient response was rescued by the cytokine IL-17A. The identification of IL-17A as the cytokine capable of reversing the poor response of CD8+ T cells provides information about a potential therapeutic treatment aimed at enhancing defective immunity induced by B cell depletion.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Antígenos CD20/imunologia , Linfócitos T CD8-Positivos/imunologia , Doença de Chagas/imunologia , Interleucina-17/imunologia , Animais , Anticorpos Monoclonais/imunologia , Doença de Chagas/prevenção & controle , Feminino , Injeções Intraperitoneais , Interleucina-17/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Trypanosoma cruziRESUMO
Classical conditioning (CC) with a sweet flavored is useful for experimental models of immune response. The objective this work was analyze spleen cell response on the differentiation of B-lymphocytes associated with the CC with sweet flavored in adult rats. Twelve adult male rats were divided (n = 3) in Group 1 (control); Group 2 without conditioned stimulus (WCS) received only water; Group 3 receiving only water with salt; Group 4 (experimental) with CS of sweet flavored and salty flavored afterwards. In every work group, all animals were subjected to the unconditioned stimulus (US) treated by 10 mg/kg body weight of cyclophosphamide injected intraperitoneally (i.p.). Subsequently an immunological challenge with sheep red blood cells i.p. was applied. Spleen samples were obtained 15 days after using euthanasia with 1% sodium thiopental i.p. The evaluation B lymphocyte (immature and mature) was performed by immunohistochemistry using specific antibodies against CD20 and against CD37, and revealed with HRP/DAB. The results show in every work group significant increases of CD20+ and CD37+ cell densities in the red pulp of the spleen of male adult rats from different groups of the experimental design. We concluded that the immunomodulatory response under CC might facilitate a less aggressive and more physiological immunological response against immunosuppression. RESEARCH HIGHLIGHTS: The immune response can be reinforced by classical conditioning. The sweet taste allows to positively condition the immune response. B lymphocytes actively participate in the immune response and classical conditioning.
Assuntos
Linfócitos B/citologia , Animais , Antígenos CD/imunologia , Antígenos CD20/imunologia , Antígenos de Neoplasias/imunologia , Linfócitos B/imunologia , Diferenciação Celular , Eritrócitos/citologia , Masculino , Ratos , Ratos Sprague-Dawley , Ovinos , Baço/citologia , Baço/imunologia , Tetraspaninas/imunologiaRESUMO
INTRODUCTION: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by an inflammatory process, with a global prevalence ranging from 0.3% to 1%. The overall cost of RA drugs is estimated in $20 billion worldwide and projected to grow to $36 billion by 2021. The current RA treatment strategy consists of the aggressive therapy directed to specific targets, after diagnostic confirmation and the stepped therapy directed by the stage of the disease, aiming at the clinical remission. Conventional (methotrexate, sulfasalazine, leflunomide) and biological (infliximab, adalimumab, tocilizumab) disease-modifying antirheumatic drugs may fail, produce only partial responses, or unwanted side effects, and consequently new antirheumatic drugs are being developed to overcome these limitations. AREAS COVERED: In this review, the authors described the technological trends and the main players involved in the R&D process related to biological compounds employed in the treatment of RA, using patent documents as a source of technological information. EXPERT OPINION: Current treatments for RA still mainly target the immune system, different inflammatory targets, and mediators. Other types of therapies have also been developed, such as vaccines and gene therapies. Despite these new techniques, the main compounds of interest remain the antibodies anti-TNF-α and anti-CD20, with novelties regarding preparation methods and combination targets.
Assuntos
Antirreumáticos/farmacologia , Artrite Reumatoide/tratamento farmacológico , Desenho de Fármacos , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais/farmacologia , Antígenos CD20/imunologia , Antirreumáticos/efeitos adversos , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Fatores Biológicos/efeitos adversos , Fatores Biológicos/farmacologia , Humanos , Patentes como Assunto , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Benign lymphoadenosis of oral mucosa (BLOM) is a common oral mucosa disease and may be regarded as a precancerous lesion. However, the association between its biological behavior and lymphocyte distribution remains unclear. Therefore, to investigate the characteristics of BLOM, we studied the infiltration of lymphocytes associated with it. The expression levels of CD74, CD20, CD3, and CD45RO were evaluated by immunohistochemical staining in 14 sam-ples from BLOM, 9 samples from BLOM with atypia hyperplasia, 11 samples from BLOM with canceration, and 10 samples from normal oral mucosa tissues. The results were analyzed by two-sample t-test using SPSS 10.0 for Windows, and P < 0.05 was considered to be sig-nificant. In normal oral mucosa, positive expression levels of CD3 and CD45RO were presented in the extra-lymphoid follicle, and the expres-sion levels of CD74 and CD20 were negative. In all BLOM groups, the expression level of CD20 was positive except for one case of BLOM with canceration; the expression levels of CD74 were all positive. Posi-tive expression levels of CD3 and CD45RO could be found not only in extra-lymphoid follicles but also in inner-lymphoid follicles in the BLOM groups. The expression levels of CD74 and CD20 in extra-lym-phoid follicles, and CD3 and CD45RO in inner-lymphoid follicles in BLOM were significantly higher than in BLOM with canceration. The infiltrated lymphocytes in BLOM comprise T- and B-cells. This indi-cates that the lymphoid tissue in BLOM is mucosa-associated lymphoid tissue and BLOM is a proliferative lesion.
Assuntos
Linfócitos B/imunologia , Expressão Gênica/imunologia , Linfoma de Zona Marginal Tipo Células B/imunologia , Mucosa Bucal/imunologia , Neoplasias/imunologia , Linfócitos T/imunologia , Antígenos CD20/genética , Antígenos CD20/imunologia , Antígenos de Diferenciação de Linfócitos B/genética , Antígenos de Diferenciação de Linfócitos B/imunologia , Linfócitos B/patologia , Complexo CD3/genética , Complexo CD3/imunologia , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Imuno-Histoquímica , Antígenos Comuns de Leucócito/genética , Antígenos Comuns de Leucócito/imunologia , Tecido Linfoide/imunologia , Tecido Linfoide/patologia , Linfoma de Zona Marginal Tipo Células B/genética , Linfoma de Zona Marginal Tipo Células B/patologia , Mucosa Bucal/patologia , Neoplasias/genética , Neoplasias/patologia , Linfócitos T/patologiaRESUMO
Rituximab is a chimeric monoclonal antibody employed for the treatment of CD20-positive B-cell non-Hodgkin's lymphoma, chronic lymphocytic leukemia, rheumatoid arthritis, granulomatosis with polyangiitis and microscopic polyangiitis. It binds specifically to the CD20 antigen expressed on pre-B and consequently on mature B-lymphocytes of both normal and malignant cells, inhibiting their proliferation through apoptosis, CDC, and ADCC mechanisms. The immunomodulatory activity of rituximab is closely related to critical quality attributes that characterize its chemical composition and spatial configuration, which determine the recognition of CD20 and the binding to receptors or factors involved in its effector functions, while regulating the potential immunogenic response. Herein, we present a physicochemical and biological characterization followed by a pharmacodynamics and immunogenicity study to demonstrate comparability between two products containing rituximab. The physicochemical and biological characterization revealed that both products fit within the same response intervals exhibiting the same degree of variability. With regard to clinical response, both products depleted CD20+ B-cells until posttreatment recovery and no meaningful differences were found in their pharmacodynamic profiles. The evaluation of anti-chimeric antibodies did not show differential immunogenicity among products. Overall, these data confirm that similarity of critical quality attributes results in a comparable immunomodulatory activity.
Assuntos
Antineoplásicos/farmacologia , Linfócitos B/imunologia , Fatores Imunológicos/farmacologia , Rituximab/farmacologia , Sequência de Aminoácidos , Antígenos CD20/imunologia , Antineoplásicos/química , Antineoplásicos/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/metabolismo , Depleção Linfocítica , Ligação Proteica/fisiologia , Rituximab/química , Rituximab/metabolismoRESUMO
Five patients with active disseminated vitiligo were given 1g of a chimeric (murine/human) monoclonal antibody to CD20 in a single intravenous infusion and followed-up for 6 months. Three of the patients showed an overt clinical and histological improvement of the disease, one presented slight improvement and the remaining patient showed no changes. Improvement was neither associated with changes in laboratory parameters nor to a specific human leucocyte antigen D-related (HLA-DR) phenotype. We believe that these preliminary results are encouraging, and further clinical trials should be undertaken. An important aim should be the finding of a marker with a good response to this therapeutic approach.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antígenos CD20/imunologia , Vitiligo/terapia , Animais , Anticorpos Monoclonais/administração & dosagem , Biomarcadores Farmacológicos/sangue , Progressão da Doença , Seguimentos , Antígenos HLA-DR/metabolismo , Humanos , Infusões Intravenosas , Camundongos , Projetos Piloto , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/uso terapêutico , Equilíbrio Th1-Th2 , Resultado do TratamentoRESUMO
Disseminated leishmaniasis (DL) differs from other clinical forms of the disease due to the presence of many non-ulcerated lesions (papules and nodules) in non-contiguous areas of the body. We describe the histopathology of DL non-ulcerated lesions and the presence of CD4-, CD20-, CD68-, CD31- and von Willebrand factor (vW)-positive cells in the inflamed area. We analysed eighteen biopsies from non-ulcerated lesions and quantified the inflamed areas and the expression of CD4, CD20, CD68, CD31 and vW using Image-Pro software (Media Cybernetics). Diffuse lymphoplasmacytic perivascular infiltrates were found in dermal skin. Inflammation was observed in 3-73% of the total biopsy area and showed a significant linear correlation with the number of vW+ vessels. The most common cells were CD68+ macrophages, CD20+ B-cells and CD4+ T-cells. A significant linear correlation between CD4+ and CD20+ cells and the size of the inflamed area was also found. Our findings show chronic inflammation in all DL non-ulcerated lesions predominantly formed by macrophages, plasmacytes and T and B-cells. As the inflamed area expanded, the number of granulomas and extent of the vascular framework increased. Thus, we demonstrate that vessels may have an important role in the clinical evolution of DL lesions.
Assuntos
Inflamação/imunologia , Leishmaniose Cutânea/imunologia , Adolescente , Adulto , Antígenos CD/imunologia , Antígenos CD20/imunologia , Antígenos de Diferenciação Mielomonocítica/imunologia , Linfócitos B/imunologia , Linfócitos B/patologia , Biópsia , Linfócitos T CD4-Positivos/imunologia , Criança , Pré-Escolar , Doença Crônica , Progressão da Doença , Feminino , Humanos , Inflamação/patologia , Leishmaniose Cutânea/patologia , Macrófagos/imunologia , Macrófagos/patologia , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Fator de von Willebrand/imunologiaRESUMO
The CD20 molecule is a non-glycosylated protein expressed mainly on the surface of B lymphocytes. In some pathogenic B cells, it shows an increased expression, thus becoming an attractive target for diagnosis and therapy. Rituximab is a chimeric antibody that specifically recognizes the human CD20 molecule. This antibody is indicated for the treatment of non-Hodgkin lymphomas and autoimmune diseases, such as rheumatoid arthritis and systemic lupus erythematosus. In this work, we describe the stable expression and biological evaluation of an anti-CD20 biosimilar antibody. While rituximab is produced in fed-batch culture of recombinant Chinese hamster ovary (CHO) cells, our biosimilar antibody expression process consists of continuous culture of recombinant murine NS0 myeloma cells. The ability of the purified biosimilar antibody to recognize the CD20 molecule on human tumor cell lines, as well as on peripheral blood mononuclear cells from humans and primates, was demonstrated by flow cytometry. The biosimilar antibody induced complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity and apoptosis on human cell lines with high expression of CD20. In addition, this antibody depleted CD20-positive B lymphocytes from peripheral blood in monkeys. These results indicate that the biological properties of the biosimilar antibody compare favorably with those of the innovator product, and that it should be evaluated in future clinical trials.
Assuntos
Anticorpos Monoclonais Murinos/imunologia , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos/imunologia , Antígenos CD20/imunologia , Animais , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais Murinos/genética , Anticorpos Monoclonais Murinos/farmacologia , Afinidade de Anticorpos/imunologia , Citotoxicidade Celular Dependente de Anticorpos/efeitos dos fármacos , Citotoxicidade Celular Dependente de Anticorpos/imunologia , Antígenos CD20/genética , Antígenos CD20/metabolismo , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Linfócitos B/metabolismo , Células CHO , Linhagem Celular Tumoral , Células Cultivadas , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/metabolismo , Cricetinae , Cricetulus , Citotoxicidade Imunológica/efeitos dos fármacos , Citotoxicidade Imunológica/imunologia , Citometria de Fluxo , Humanos , Células K562 , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Macaca fascicularis , Engenharia de Proteínas/métodos , RituximabRESUMO
Classification of acute lymphoblastic leukemia (ALL) by flow cytometric immunophenotyping characterizes the disease and delineates potential therapeutic intervention. We retrospectively analyzed CD20 expression in 143 patients with newly diagnosed precursor B-cell ALL. CD20 was observed in 61% of patients at diagnosis. There was no correlation between CD20 expression and age, white blood cell count, or cytogenetic abnormalities. Despite the fact that CD20-positive ALL patients had a tendency toward a worse outcome, there was no significant difference between patients with and without CD20 expression in 3-year overall survival 65 vs. 82% (P = 0.14), and cumulative incidence of relapse 36 vs. 18% (P = 0.3) in pediatric patients and 51 vs. 53% (P = 0.31) and 35 vs. 38% (P = 0.6) in adults, respectively. In conclusion, CD20 expression appears to be more common in Mexican patients with newly diagnosed precursor B-cell ALL higher than in Caucasian populations and lacks prognostic value.
Assuntos
Antígenos CD20/análise , Linfócitos B/patologia , Biomarcadores Tumorais/análise , Leucemia-Linfoma Linfoblástico de Células Precursoras B/diagnóstico , Doença Aguda , Adolescente , Adulto , Idoso , Antígenos CD20/genética , Antígenos CD20/imunologia , Linfócitos B/imunologia , Biomarcadores Tumorais/imunologia , Criança , Pré-Escolar , Intervalo Livre de Doença , Feminino , Citometria de Fluxo , Expressão Gênica/imunologia , Humanos , Imunofenotipagem , Lactente , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Leucemia-Linfoma Linfoblástico de Células Precursoras B/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/mortalidade , Valor Preditivo dos Testes , Recidiva , Estudos RetrospectivosRESUMO
Eight patients with refractory lupus nephritis received rituximab after failing standard sequential therapy and were followed for 104 weeks after the infusion. One patient died secondary to a complicated pregnancy but had stable renal function. Three patients received a re-infusion of rituximab approximately 12 months apart due to a renal flare; during the second year of follow-up, those patients progressed toward ESRD. The four remaining patients demonstrated improvements in SLEDAI score, CrCl, and proteinuria with maintenance of their standard immunosuppressive therapy and did not require a re-infusion of rituximab. Although rituximab as induction therapy for refractory lupus nephritis has been shown to have a good response, its efficacy in long-term assessments demonstrates disappointing results.