BGB-A445, a novel non-ligand-blocking agonistic anti-OX40 antibody, exhibits superior immune activation and antitumor effects in preclinical models / 医学前沿

OX40 is a costimulatory receptor that is expressed primarily on activated CD4+, CD8+, and regulatory T cells. The ligation of OX40 to its sole ligand OX40L potentiates T cell expansion, differentiation, and activation and also promotes dendritic cells to mature to enhance their cytokine production. Therefore, the use of agonistic anti-OX40 antibodies for cancer immunotherapy has gained great interest. However, most of the agonistic anti-OX40 antibodies in the clinic are OX40L-competitive and show limited efficacy. Here, we discovered that BGB-A445, a non-ligand-competitive agonistic anti-OX40 antibody currently under clinical investigation, induced optimal T cell activation without impairing dendritic cell function. In addition, BGB-A445 dose-dependently and significantly depleted regulatory T cells in vitro and in vivo via antibody-dependent cellular cytotoxicity. In the MC38 syngeneic model established in humanized OX40 knock-in mice, BGB-A445 demonstrated robust and dose-dependent antitumor efficacy, whereas the ligand-competitive anti-OX40 antibody showed antitumor efficacy characterized by a hook effect. Furthermore, BGB-A445 demonstrated a strong combination antitumor effect with an anti-PD-1 antibody. Taken together, our findings show that BGB-A445, which does not block OX40-OX40L interaction in contrast to clinical-stage anti-OX40 antibodies, shows superior immune-stimulating effects and antitumor efficacy and thus warrants further clinical investigation.

Avaliação do perfil de expressão das moléculas OX40, 4-1BB e CD40/CD40L no mieloma múltiplo / Evaluation of the expression profile of the molecules OX40, 4-1BB and CD40/CD40L in multiple myeloma

Introdução: O mieloma múltiplo é uma desordem clonal das células plasmocitárias e, responde por 10-15% das neoplasias hematológicas. Apresenta diversas alterações no sistema imune, caracterizadas por déficits na produção de anticorpos; alterações do perfil imunológico das células T; aumento da expressão do PD-L1; modificações no microambiente medular favorecendo o recrutamento de populações imunossupressoras como as Treg e disfunção nas células dendríticas. Manter um sistema imune ativo é fundamental para o controle da doença, pacientes com manutenção de células T efetoras apresentam maiores taxas de remissão e sobrevida. Receptores coestimuladores como o OX40, CD40/CD40L e 4-1BB, participam na ativação, proliferação e amplificação da resposta imune. Objetivo: Avaliar os níveis de linfócitos B e T e das moléculas coestimuladoras OX40, CD40, CD40L e 4-1BB no sangue e medula óssea dos pacientes com mieloma múltiplo. Métodos: Trata-se de estudo exploratório, realizado entre 2016 e 2019 no Hospital de Câncer de Pernambuco (HCP) e Laboratório de Pesquisa Translacional do Instituto de Medicina Integral Prof. Fernando Figueira (IMIP). Foram incluídas 40 pacientes, até 79 anos de idade, com diagnóstico de Mieloma Múltiplo. Coletas de sangue periférico e medula óssea foram realizadas ao diagnóstico. As mensurações dos níveis de expressão de proteínas de membrana CD20, CD3, OX40, CD40/CD40L foram detectadas pela técnica de Cytometric Bead Array por citometria de fluxo. A dosagem dos níveis solúveis de s4-1BB, sOX40 e sCD40L foi realizada por enzyme linked immunonoSorbent assay (ELISA). Foi realizada análise de curva Receiver Operating Characteristic (ROC) para determinar o melhor valor de acurácia de cada marcador estudado assim como, a ocorrência de óbito. A análise estatística foi realizada no programa GraphPadPrism v8.0. O nível de significância estatística foi de p<0,05. Resultados: Em sangue periférico, comparando-se pacientes e controles, verificou-se níveis menores de CD20 (p<0,0001) e CD20low (p<0,0001), CD40+ em leucócitos totais (p=0,0005), CD40+ em linfócitos (0,0006) e CD40/CD3+ (p<0,0001) no grupo de pacientes. Mas, em contrapartida, os pacientes apresentaram níveis mais elevados de OX40+ (p=0,0012), CD40L+ em leucócitos totais (p=0,002) e OX40+/CD3+ (p<0,0001). Os níveis séricos de s4-1BB (p=0,03) e sOX40 (p=0,01) estavam reduzidos no grupo de MM quando comparado aos controles. Na análise segundo o ISS, somente os níveis de expressão de CD40L+ em leucócitos (p=0,01) e de CD40+ em linfócitos (p=0,0045), mostraram níveis superiores nos pacientes com ISS1-2 em relação ao ISS-3. As medidas de expressão de OX40+ e CD40L+ em leucócitos totais eram inferiores nos casos com evolução para óbito (p<0,0006 e p=0,002, respectivamente). Os pacientes que apresentavam níveis de expressão de OX40 em leucócitos totais ≥2,93% tiveram maior sobrevida em relação àqueles com valores <2,93% (p=0,03), bem como aqueles com CD40L em leucócitos totais com valores ≥3,09% (p=0,001). Na análise da medula óssea, segundo o ISS, somente os níveis de expressão de OX40/CD3+, mostraram níveis superiores nos pacientes com ISS1-2 em relação ao ISS-3 (p<0,0017). Não foram observadas diferenças significativas entre os valores de expressão dos diversos marcadores em medula óssea, com relação ao desfecho óbito. Na análise de correlação de Spearman, os valores de CD20 em sangue e medula óssea, apresentam uma correlação moderada entre si (r=0,64 e p<0,0001). Conclusão: Os resultados deste estudo permitem concluir que existem alterações de mecanismos celulares envolvidos na regulação e ativação da resposta imune no MM quando comparados aos controles. A manutenção de níveis mais elevados de moléculas coestimuladoras (OX40 ≥2,93% e CD40L≥ 3,09%), foi preditiva de melhor sobrevida no MM

Introduction: Multiple myeloma (MM) is a malignant plasma cell (PC) disorder, accounting for approximately 10-15% of all hematological cancers. Multiple myeloma presents several immune system alterations, characterized by deficits in antibody production, disruption of the T-cell immune profile, increased expression of cell death ligand 1 (PD-L1), changes in the bone marrow microenvironment favoring the recruitment of immunosuppressive populations such as Tregs and dysfunction in dendritic cells. It is important to preserve the integrity of the active immune system for the control of disease progression and patients with maintenance of T-cell cytotoxic activities improve rates of remission and overall survival. Co-stimulating receptors such as OX40, CD40/CD40L and 4-1BB, cooperate in the activation, proliferation, and amplification of the immune response. Objective: To evaluate T and B lymphocyte levels as well as co-stimulating molecules OX40, CD40, CD40L and 4-1BB in the blood and bone marrow of multiple myeloma patients. Methods: This is a cross-sectional and exploratory study, conducted between 2016 and 2019 at Pernambuco Cancer Hospital (HCP) and Translational Research Laboratory of Instituto de Medicina Integral Prof. Fernando Figueira (IMIP). Forty patients, up to 79 years of age, diagnosed with Multiple Myeloma were included. Peripheral blood and bone marrow samples were collected at diagnosis. Serum concentrations of CD20, CD3, OX40, CD40/CD40L were detected through the Cytometric Bead Array technique by flow cytometry, and the soluble forms of s4-1BB, sOX40 e sCD40L by enzyme linked immunosorbent assays. Receiver Operating Characteristic (ROC) curve analysis was performed to determine not only the best accuracy value of each studied marker but also, mortality. Statistical analysis was performed in the GraphPadPrism v8.0 program and the level of statistical significance was p <0.05. Results: In peripheral blood, comparing patients and controls, there were lower levels of CD20 (p<0.0001) and CD20low (p<0.0001), CD40+ in total leukocytes (p=0.0005), CD40+ in lymphocytes (0.0006) and CD40/CD3+ (p<0.0001) in the patient group. However, on the other hand, patients had higher levels of OX40+ (p=0.0012), CD40L+ in total leukocytes (p=0.002) and OX40+/CD3+ (p<0.0001). Serum levels of s4-1BB (p=0.03) and sOX40 (p=0.01) were reduced in the MM group compared to controls. According to the ISS, CD40L+ in leukocytes (p=0.01) and CD40+ in lymphocytes (p=0.0045) showed higher levels in patients with ISS1-2 compared to ISS-3. Regarding the outcome death, levels of OX40+ and CD40L+ in total leukocytes were lower (p<0.0006 and p=0.002, respectively). In survival analyses, patients who had OX40+ levels in total leukocytes ≥2.93% had higher survival compared to those with levels <2.93% (p=0.03), as well as those with CD40L+ in total leukocytes with values ≥3.09% (p=0.001). In the bone marrow only the OX40/CD3+ levels were higher in patients with ISS1-2 compared to ISS-3 (p<0.0017). No significant differences were observed between values of other bone marrow markers in relation to the outcome death. In Spearman's correlation analysis, CD20 levels in blood and bone marrow present moderate correlation between them (r=0.64 and p<0.0001). Conclusion: This study shows differences in cellular mechanisms involved in the regulation and activation of immune response in MM patients in comparison to healthy controls. The maintenance of higher levels of co-stimulating molecules (OX40 ≥2.93% and CD40L≥ 3.09%) is associated with better survival in multiple myeloma

Análise da expressão de microRNAs, receptores OX40 e CD40L, e do agregado de plaquetas-leucócitos no câncer gástrico / Analysis of the expression of microRNAs, OX40 and CD40L receptors, and platelets-leukocytes aggregation in gastric cancer

O prognóstico dos portadores de câncer gástrico tem melhorado pouco nas últimas décadas e o melhor entendimento das vias moleculares e as interações imunes no microambiente tumoral podem revelar novas possibilidades de tratamento. O ambiente tumoral é composto por células do sistema imune, que refletem a tentativa desse sistema em promover uma resposta antitumoral. Complexas interações entre células e mediadores imunes no microambiente tecidual regulam o crescimento de tumores, progressão, metástase e angiogênese. O entendimento das alterações da imunidade na população com câncer gástrico (CG) permitirá a intervenção terapêutica para melhorar a resposta à cirurgia e à quimioterapia. Objetivo: comparar a expressão de miRNA em tecidos de pacientes com câncer gástrico e controles saudáveis para encontrar miRNAs desregulados no câncer gástrico e usar ferramentas de bioinformática para determinar a possível influência desses miRNAs no sistema imunológico. Avaliar a formação de agregados plaquetas-leucócitos circulantes, bem como os níveis de ativação plaquetária (CD62p+, CD40L) desses conjugados a leucócitos em pacientes com CG. Quantificar a expressão de moléculas costimulatórias da resposta imune (OX40) em linfócitos T de pacientes com CG. Métodos: é um estudo do tipo corte transversal, exploratório e translacional realizado no Hospital de Câncer de Pernambuco, Laboratório de Pesquisa Translacional do Instituto de Medicina Integral Prof. Fernando Figueira e Centro Internacional de Pesquisa (CIPE) do AC Camargo entre 2015 a 2018. Foram avaliados 83 pacientes com câncer gástrico e 69 controles. Foram determinados os níveis de expressão de microRNAs no tecido tumoral gástrico em comparação aos da mucosa gástrica normal por técnica de reação em cadeia da polimerase (qPCR - TaqMan). Foram realizadas as análises dos niveis linfócitos T e B, OX40, CD40L e de agregado de plaquetas no sangue periférico por citometria de fluxo. Resultados: As análises revelaram um miRNA mais expresso (miR-196a-5p) e dois significativamente menos expressos (miR-374a-5p e miR-375) em comparação ao grupo controle. Pacientes com estádio IV (metástatico) apresentaram uma diminuição significativa na expressão do miR-374-5p quando comparados aos pacientes não metastáticos (p=0.03). Com uso de plataformas de bioinformática, foram observadas várias vias que sofrem influência dos microRNAs desregulados e que interagem com genes envolvidos com a resposta imune celular, moléculas de adesão celular e migração celular. Foram encontrados níveis elevados de expressão de OX40 em linfócitos T, monócitos e neutrófilos de pacientes com neoplasia gástrica (p<0.0001), entretanto, os níveis de OX40 foram reduzidos nos grupos com neoplasia estádio III/IV quando comparados ao estádio I/II. Observamos níveis elevados de agregados de plaquetas-linfócitos T e plaquetas-linfócitos B no sangue de pacientes GC com estágio IV quando comparados com os estágios I, II e III, e grupo controle (p <0,05). Níveis reduzidos de agregados plaquetas-linfócitos totais com expressão de CD40L foram observados no estádio IV da doença (p<0,05). Níveis elevados de plaquetas ativadas e agregados de plaquetas-monócitos ativados (CD62p+) foram observados em pacientes GC quando comparados ao grupo controle (p<0,05). Conclusão: Os resultados deste estudo permitem concluir que existem alterações de mecanismos moleculares (miRNA) e celulares envolvidos na regulação e ativação da resposta imune, sendo associadas à progressão e metástase no GC

The prognosis of gastric cancer patients has not been improved in the last decades and the the understanding of the molecular immune pathways and how immune interactions happens on tumoral microenvironment, open new possibilities of treatment. The tumor environment is infiltrated by cells of the immune system, which reflect an antitumor response. Complex interactions between microenvironmental cells and mediators regulate tumor growth, progression, metastasis and angiogenesis. The knowledge of the immunity in the population with gastric cancer will allow therapeutic intervention to improve the response to surgery and the chemotherapy. Objective: our primary goal was to compare the miRNA expression in tissues from primary gastric cancer patients and healthy controls to find miRNAs dysregulated in gastric cancer and used bioinformatics tools to determine potential roles of these miRNAs in the immune system. We conducted a secondary analysis to evaluate the formation of circulating platelet-leucocyte conjugates as well as the CD40L levels conjugate to leucocytes in GC and finally, samples were analysed for levels of costimulatory molecules related to the immune response (OX40) in GC. Methods: A cross-sectional, translational and exploratory study carried out at the Hospital de Cancer de Pernambuco, Translational Research Laboratory of the Instituto de Medicina Integral Prof. Fernando Figueira and AC Camargo International Research Center (CIPE) from 2015 to 2018, involving 83 patients with gastric cancer and 69 controls. Expression levels of microRNAs in gastric tumor tissue and normal gastric mucosa were determined by polymerase chain reaction technique (qPCR - TaqMan). Analyzes of T and B lymphocytes, OX40, CD40L and platelet aggregate levels were performed in peripheral blood by flow cytometry. Results: The results revealed a more express miRNA (miR-196a-5p) and two significantly less expressed (miR-374a-5p and miR-375) compared to the control group. Patients with stage IV (metastasis) showed a significant decrease in miR-374-5p expression when compared to non-metastatic patients (p = 0.03). Bioinformatics analysis suggested that the pathways regulated by these differentially expressed miRNAs were related to the immune response, cell adhesion, and cell migration. High levels of OX40 expression were found in T lymphocytes, monocytes and neutrophils of patients with gastric neoplasia (p <0.0001); however, OX40 levels were reduced in groups with stage III / IV neoplasia when compared to stage I / II. We observed higher levels of platelet-T lymphocyte aggregate (P-T lymp) and platelet-B lymphocyte aggregate (P-B lymp) in the peripheral blood (PB) of GC patients with stage IV when compare with stages I-II-III, and control group (p<0,05). Reduced levels of CD40L+ Platelet-total lymphocyte (P-lymp) were observed at stage IV of the disease (p<0.05). High levels of CD62p+ platelets and CD62p+ platelets-monocyte aggregate were observed GC patients when compare to control group (p<0.05). Conclusion: The results of this study allow us to conclude that there are alterations of molecular mechanisms (miRNA) and cellular mechanisms involved in the regulation and activation of the immune response and associated to the progression and metastasis in GC

Aberrant Expressions of Immune Factors Facilitate the Disequilibrium of Immune Status in Cervical Cancer / 中国医学科学院学报

Objective To explore the expressions and co-relationship of immune factors forkhead box p3 (FoxP3),chemokine (C-C motif) ligand 22 (CCL22),tumor necrosis factor receptor superfamily member 40(OX40),and SMAD family member 3 (Smad3) in cervical carcinoma and investigate their immunomodulatory roles in cervical carcinogenesis.Methods Totally 30 cases of cervical carcinoma with adjacent tissues and 20 cases of normal cervix were collected in this study. FoxP3,CCL22,OX40,and Smad3 mRNA expressions were detected by real-time polymerase chain reaction (RT-PCR). Results Compared to normal cervix,the expression levels of FoxP3 and CCL22 mRNA were elevated in neoplastic foci(P=0.000,P=0.002) and tumor periphery (P=0.048,P=0.040).The mRNAs increased modestly in high-grade squamous cell carcinoma focal(P=0.019,P=0.020) and periphery tissue (P=0.023,P=0.031) in comparison with low-grade squamous cell carcinoma. The expression levels of OX40 and Smad3 mRNA were significantly lower in neoplastic foci(P=0.000,P=0.015) than normal cervix. Compared to low-grade squamous cell carcinoma focal and periphery tissue,the mRNAs decreased moderately in high-grade squamous cell carcinoma(P=0.018,P=0.030; P=0.027,P=0.014). In both neoplastic foci and tumor periphery,the mRNA expression level of CCL22 was positively correlated with FoxP3 (r=0.353,P=0.000; r=0.307,P=0.000) but negatively correlated with OX40 (r=-0.288,P=0.031; r=-0.263,P=0.037),while OX40 was positively correlated with Smad3 (r=0.384,P=0.002;r=0.288,P=0.023). The mRNA expressions of FoxP3 and CCL22 were increased in foci and pericarcinous tissues (P=0.024,P=0.039; P=0.032,P=0.034) while Smad3 was decreased in neoplastic foci (P=0.017) in contrast to HPV negative corresponding group. Conclusion FoxP3 and CCL22 expressions increase while OX40 and Smad3 expression decrease at mRNA level in the microenvironment of cervical cancer,which may be associated with such immunological model that the immunosuppressive roles of FoxP3 and CCL22 enhance while the immunity-boosting roles of OX40 and Smad3 are impeded,contributing to the deterioration of immune disequilibrium in local site and cervical cancer carcinogenesis.

Blockade of the OX40/OX40L pathway and induction of PD-L1 synergistically protects mouse islet allografts from rejection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>OX40/OX40 ligand (OX40/OX40L) and programmed death-1/programmed death ligand-1 (PD-1/PD-L1) costimulatory signals play important roles in T cell-induced immune responses. The aim of this study was to investigate the roles of OX40/OX40L and PD-1/PD-L1 costimulatory pathways in mouse islet allograft rejection.</p><p><b>METHODS</b>Lentiviral vectors containing OX40L siRNA sequences and an adenovirus vector containing the PD-L1 gene were constructed. The streptozotocin-induced model of diabetes was established in C57BL/6 (H-2(b)) mice. Diabetic C57BL/6 mice were randomly allocated into five groups: group 1, untreated control; group 2, Ad-EGFP treatment; group 3, Ad-PD-L1 treatment; group 4, OX40L-RNAi-LV treatment; group 5, OX40L-RNAi-LV combined with Ad-PD-L1 treatment. Lentiviral vector and the adenovirus vector were injected, singly or combined, into the caudal vein one day before islet transplantation. The islets of DBA/2 (H-2(d)) mice were transplanted into the renal subcapsular space of the diabetic recipients. Recipient blood glucose and the survival time of the allografts were monitored. Antigen-specific mixed lymphocyte reaction was also evaluated.</p><p><b>RESULTS</b>The recombinant lentiviral RNA interference vector OX40L-RNAi-LV reduced OX40L protein expression by 70%. The recombinant adenovirus vector Ad-PD-L1 increased PD-L1 protein expression in vivo in C57BL/6 recipient mice. Combined OX40L-RNAi-LV/Ad-PD-L1 treatment induced a synergistic protective effect in pancreatic islet allografts. Allograft survival time in the combined treatment group was (92.27±9.65) days, not only longer than that of the control ((6.51±0.27) days) and Ad-EGFP groups ((7.09±0.13) days) (P < 0.01), but also significantly longer than that of Ad-PD-L1 and OX40L-RNAi-LV single treatment groups ((40.64±3.95) days and (55.14±5.48) days respectively, P < 0.01). The blood glucose concentration of recipient mice in the combined treatment group was also stable and kept within the normal range. Flow cytometry analysis showed that combined OX40L-RNAi-LV/Ad-PD-L1 treatment significantly decreased proliferation in an antigen-specific mixed lymphocyte reaction. After donor DBA/2 lymphocyte stimulation, 89.71% of lymphocytes from recipient combination treatment C57BL/6 mice were not split and proliferated. In contrast, after stimulation with third party Lewis rat lymphocytes, only 45.84% lymphocytes of C57BL/6 mice were not split and proliferated.</p><p><b>CONCLUSIONS</b>This study demonstrates the successful construction of the recombinant lentivirus vector OX40L-RNAi-LV and adenovirus vector Ad-PD-L1 for the blockade of OX40/OX40L and activation of PD-1/PD-L1 costimulatory pathways simultaneously in pancreatic islet allografts in diabetic mice. Combination therapy with these two vectors resulted in inhibition of T cell activation, synergistically prolonging the survival time of pancreatic islet allografts.</p>

Concomitant increase of OX40 and FOXP3 transcripts in peripheral blood of patients with breast cancer

Regulatory T cells [T-regs] have an important role in cancer by suppression of protective antitumor immune responses. Regulatory T cells express the forkhead/ winged helix transcription factor [FOXP3] and OX40 molecules which have important regulatory roles in the immune system. To evaluate FOXP3 and OX40 transcripts in the peripheral blood mononuclear cells of women with breast cancer. Blood samples from 40 women with histologically-confirmed infiltrating ductal carcinoma of the breast and 40 healthy volunteer women without a history of malignancy or autoimmune disorders were collected. The abundance of FOXP3 and OX40 gene transcripts were determined by quantitative real-time PCR [qRT-PCR]. There was a significant positive correlation between FOXP3 and OX40 gene expression in women with breast cancer in a stage dependent manner. This finding emphasizes the importance of T-regs as predominant targets for breast cancer immunotherapy

OX40 mRNA in peripheral blood as a biomarker of acute renal allograft rejection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Acute rejection remains an important cause of renal allograft dysfunction and the need for accurate diagnosis is essential to successfully treat transplant recipients. The purpose of this study was to determine the costimulatory molecules OX40 and OX40L messenger RNA (mRNA) levels in peripheral blood mononuclear cells (PBMCs) to predict acute renal transplant rejection.</p><p><b>METHODS</b>The whole blood samples from 20 recipients with biopsy-confirmed acute rejection (rejection group), 20 recipients with stable graft function and normal biopsy results (stable group) after kidney transplantation, and 20 healthy volunteers (control group) were collected. The mRNA levels of OX40 and OX40L were analyzed with TaqMan real-time reverse transcriptase polymerase chain reaction (RT-PCR). The association of OX40 and OX40L mRNA levels with disease severity was investigated.</p><p><b>RESULTS</b>There was no significant difference of OX40, OX40L mRNA levels in PBMCs between the stable group and control group (P > 0.05). The levels of OX40 and OX40L mRNA were significantly higher in the rejection group than in the control group (P < 0.01 and P < 0.05, respectively). Non-significantly higher OX40L mRNA and significantly higher OX40 mRNA in PBMCs were observed in subjects in the rejection group compared with the stable group (P > 0.05 and P < 0.01, respectively). Receiver operating characteristic (ROC) curve analysis demonstrated that OX40 mRNA levels could discriminate recipients who subsequently suffered acute allograft rejection (area under the curve, 0.908). OX40 and OX40L mRNA levels did not significantly correlate with serum creatinine levels in the rejection group (P > 0.05). Levels of OX40 mRNA after anti-rejection therapy were lower than those at the time of protocol biopsy in the rejection group (P < 0.05).</p><p><b>CONCLUSION</b>Our data suggest that measurement of OX40 mRNA levels after transplant might offer a noninvasive means for recognizing recipients at risk of acute renal allograft rejection.</p>

Effect of the costimulatory molecules OX40-OX40 ligand interaction on the expression of NFATc1 in leukocytes of apolipoprotein E-deficient mice / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effect of OX40/OX40L interaction on the nuclear factor of activated T cells c1 (NFATc1) in ApoE-/- mice.</p><p><b>METHODS</b>Lymphocytes were prepared from mouse spleens after Collar-treated Surgery, then incubated with a range of agonistic anti-OX40 mAbs and inhibitory anti-OX40L mAb to stimulate or inhibit OX40-OX40L interaction in vitro. The expression of NFATc1 mRNA and protein in lymphocytes of ApoE-/- mice was measured by Real Time PCR and flow cytometry, respectively.</p><p><b>RESULTS</b>(1) After stimulating OX40-OX40L signal pathway, the expression of NFATc1 mRNA and protein in leukocytes of ApoE-/- mice was significantly increased, with maximal effect occurring at 20 µg/ml anti-OX40 mAb-stimulated, and peaked at 24 h at any concentration (P < 0.01). (2) Anti-OX40L mAb significantly suppressed the expression of NFATc1 in leukocytes of ApoE-/- mice, with maximal effect occurring at 20 µg/ml anti-OX40L mAb, and peaked at 24 h (P < 0.001).</p><p><b>CONCLUSIONS</b>OX40-OX40L interaction can regulate the mRNA and protein expressions of NFATc1 in lymphocytes of ApoE-/- mice.</p>

OX40/OX40L in systemic lupus erythematosus: association with disease activity and lupus nephritis

OX40-OX40L interaction is implicated in the pathogenesis of systemic lupus erythematosus [SLE]. We evaluated the role of OX40/OX40L as markers of disease activity and nephritis in SLE patients. Case-control study conducted in 2009 on SLE patients attending the outpatient clinics of Ain Shams University Hospital, Egypt. We assessed the percentage of CD4+ T-lymphocytes expressing OX40 by flowcytometry, and serum OX40 ligand [OX40L] levels in 40 patients with SLE [20 with lupus nephritis and 20 without] and in 20 healthy controls. Disease activity was assessed by the University of Toronto SLE disease activity index [SLEDAI]. The percentage of CD4+ T-lymphocytes expressing OX40 was significantly higher in SLE patients than in controls, and in patients with lupus nephritis than in those without. OX40 expression correlated positively with both serum creatinine levels and SLEDAI. OX40 expression was the highest in patients with class V lupus nephritis and lowest in class II. Serum OX40L levels were significantly higher in SLE patients than in controls, and in patients with nephritis than in those without. Serum OX40L levels correlated with serum creatinine levels but not with SLEDAI. OX40 expression on CD4+ T-cells had a higher sensitivity and specificity in diagnosing lupus nephritis than both OX40L and anti-double-stranded DNA levels. OX40-OX40L interaction plays a role in the pathogenesis of SLE. The expression of OX40 on CD4+ T-lymphocytes and the serum level of OX40L may act as markers of lupus nephritis. Measurements of percentages of CD4+ T-lymphocytes expressing OX40 may serve as an indicator of disease activity in SLE

Association of rs2298212 polymorphism in OX40 gene with coronary atherosclerotic disease in Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the association of the OX40 gene rs2298212G/A polymorphism with coronary atherosclerotic disease (CAD) in Chinese Han population.</p><p><b>METHODS</b>Five hundred and thirty six CAD patients and 544 age and ethnic matched controls of Chinese Han population were recruited from Qilu Hospital, Shandong University. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to genotype the selected single nucleotide polymorphism. Distributions of genotypic and allelic frequencies were analyzed by Chi-square test.</p><p><b>RESULTS</b>The distribution of genotypic and allelic frequencies have no significant differences between the CAD cases and controls(P> 0.05), even after adjusting for age, gender, body mass index, systolic blood pressure, diastolic blood pressure, glucose, total cholesterol, and triglyceride. However, when substratification analysis of the involved coronary artery vessels was performed, significant difference was found between single-vessel and triple-vessel (P= 0.02, OR = 1.56, 95% CI: 1.08-2.26) involvement.</p><p><b>CONCLUSION</b>The rs2298212G/A polymorphism in OX40 gene may be associated with the severity of coronary atherosclerotic disease.</p>

Dynamic change OX40/OX40L mRNA in experimental allergic neuritis / 中南大学学报(医学版)

OBJECTIVE@#To examine the expression of mRNA of Oxford 40(OX40) and Oxford 40 ligand(OX40L) in the sciatic nerve, spleen, peripheral blood mononuclear cells and lymph nodes of experimental allegic neuritis (EAN).@*METHODS@#Thirty-six Lewis rats were randomly assigned into an EAN group and a CFA group. The rats were sacrificed on 9th, 17th, and 26th day after immunization. OX40 and OX40L mRNA was detected by reverse transcription polymerase chain reaction in the spleen, sciatic nerves, peripheral blood mononuclear cells and lymphonodes.@*RESULTS@#The peak of clinical course came on 17th day after the immunization in EAN. The mRNA expression of OX40/OX40L was higher on 8th day and 17th day than that on 26th day after the immunization (P<0.05). There was significant difference between the EAN group and the CFA group at the 3 time points (P<0.05); rats in the CFA group didn't have any clinical manifestations. The mRNA expression of OX40 and OX40L in the EAN group raised in the sciatic nerves and lymph nodes at the above 3 time points (P<0.05). Weak expression was seen in the peripheral blood mononuclear cells.@*CONCLUSION@#OX40 and OX40L may play a role in the pathogenesis of experimental allegic neuritis.

Costimulatory molecules CD80 and CD134 associated with lupus nephritis, skin lesions and disease activity in systemic lupus erythematosus

Studying the role of CD80, CD134 and soluble CD134 ligand [CD134L] costimulatory molecules in SLE, and their correlation with disease activity and renal involvement. Also, correlation between skin lesions, SLE activity and costimulatory molecules expression. Forty patients with SLE and 40 apparently healthy controls were included in this the study. Twenty patients had biopsy-proven lupus nephritis. Clinical disease activity was assessed according to systemic lupus erythematosus disease index [SLEDAI]. CD4+ T cell populations in the peripheral blood were analyzed for the expression of co-stimulatory markers CD134 and CD80. CD134L in the serum of SLE patients and controls was also measured. SLE patients showed an increased soluble CD134L concentration in the serum and increased frequency of peripheral CD4+ T cells expressing high levels of CD80 and CD134 compared to healthy controls. These costimulatory molecules were significantly higher in SLE patients with lupus nephritis compared to patients without nephritis and they were significantly correlated with SLEDAI. Our study revealed also that the presence of skin lesions specific for LE was associated with a milder disease, also patients with only LE-nonspecific skin lesions showed an increased frequency of peripheral CD4+ T cells expressing high levels of CD80 and CD134 and soluble CD134L concentration in the serum compared to those with only LE-specific skin lesions. Increased expression of GD80 and CD134 on CD4+ T cells and increased serum concentration of soluble CD134L are associated with increased incidence of renal disease, disease activity and more serious skin lesions in SLE

Expression of OX40 and Bcl-2 in allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the role of OX40 and Bcl-2 in allergic rhinitis(AR).@*METHOD@#Twenty-three subjects of allergic rhinitis as experimental group were collected and 20 subjects of deflection of nasal septum as control group. OX40 and Bcl-2 expression were examined in biopsy specimens of AR patients and nasal septum patients by immunohistochemistry respectively. The relationship between OX40 and Bcl-2 expression was analyzed.@*RESULT@#OX40 and Bcl-2 expression increased in the AR patients. There were positive staining not only in CD4+ T cells,but also in vascular endothelial cell, epithelial cell and glandular epithelium cell (P < 0.01). Compared with control group, the expression of OX40 (0.239 4 +/- 0.033 5) and Bcl-2 (0.237 3 +/- 0.042 1) were significantly higher than that in control group (P < 0.01). The OX40 expression was closely related to Bcl-2 expression (r = 0.869 0, P < 0.05).@*CONCLUSION@#The abnormal expression of costimulatory molecule OX40 is well evidenced in AR. Bcl-2 expression is likely mediated by OX40 signal pathway.