[Chlorpromazine hydrochloride plays a tumor suppressive role in diffuse large B lymphoma by promoting the expression of S1PR2].

Objective: To investigate the effect of chlorpromazine hydrochloride on proliferation, apoptosis and cell cycle arrest of human diffuse large B lymphoma cells and its possible mechanism. Methods: In January 2019, OCI-LY3 and TMD8 cells were treated with different concentrations of chlorpromazine hydrochloride for 72 h, the proliferations were detected by Alamar Blue assay. The apoptosis and cell cycle arrest of OCI-LY3 and TMD8 cells were detected by flow cytometry with FITC annexin V/PI (propidium iodide) double staining and PI, respectively. The mRNA levels of cyclin-dependent kinase inhibitor P21, P27, CyclinD1 and S1PR2 were detected by RTqPCR. The protein levels of P21, P27 and S1PR2 were detected by Western blot. Results: With the increase of chlorpromazine hydrochloride concentration, the proliferation inhibition rates of OCI-LY3 and TMD8 cells increased at 72 hours, apoptosis and G1 cell cycle arrest increased at 24 hours. Compared with the control group, the expression levels of P21, P27 and SIPR2 mRNA in OCI-LY3 and TMD8 cells were increased at 10 µmol/L chlorpromazine after 12 hours treatment (P<0.05) . And there was no significant difference in the expression of CyclinD1 mRNA (P>0.05) . There was a similar increase in protein levels of P21, P27 and SIPR2 in OCI-LY3 and TMD8 cells after 24 hours of treatment (P<0.05) . Conclusion: Chlorpromazine hydrochloride at a specific concentration may inhibit the proliferation of ABC diffuse large B lymphoma cells by promoting the expression of S1PR2, and promote cell apoptosis and G1 phase cell cycle arrest.

Major histocompatibility complex-restricted lysis of neuroblastoma cells by autologous cytotoxic T lymphocytes.

Vigorous host immune reactivity to neuroblastoma may correlate with better prognosis, but identification of human cytotoxic T-lymphocyte (CTL) responses has been relatively unsuccessful. We generated neuroblastoma-reactive CTL lines from two human leukocyte antigen (HLA) A2+ neuroblastoma patients by stimulation of peripheral blood lymphocytes (PBLs) with irradiated autologous tumor cells pretreated with interferon-gamma in the presence of low concentrations of interleukin-2 (5 U/mL). These lines lyse autologous tumor cells but do not kill HLA mismatched allogeneic tumor cells, Epstein-Barr virus-transformed autologous B cells, or standard natural killer cell targets. Cytotoxic T lymphocytes generated from one patient recognize tumor cells from several HLA-A2 matched children, although the other patient's CTLs do not kill tumor cells from other HLA-A2+ individuals. Pretreatment of CTLs or target cells with appropriate standard monoclonal antibodies demonstrates that these CTLs are major histocompatibility complex class I (HLA-A2) restricted and that the effector cell population is CD8+. Our findings suggest that these tumor cells express at least one common HLA-A2 restricted antigen and at least one unique private epitope. Autologous tumor-specific CTLs can be readily generated from patients' PBLs and maintained in long-term culture using standard techniques.

T cell receptor BV gene rearrangements in the spinal cords and cerebrospinal fluid of patients with amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal disorder whose etiology and pathogenesis remain unknown. Recent studies, however, have demonstrated the presence of inflammatory infiltrates within ALS spinal cord and suggested the possibility of an immune-mediated process in motor neuron degeneration. We have analyzed the diversity of T-cells in the spinal cord in ALS. Reverse transcriptase polymerase chain reaction (RT-PCR) with variable (V) region sequence specific oligonucleotide primers was used to amplify T-cell receptor (TCR)BV transcripts from spinal cords obtained at autopsy from patients with ALS, patients who died without inflammatory disease of the central nervous system, brains from patients with ALS, and brains from patients who died with inflammatory CNS disease. Sequencing was then performed on the amplified transcripts. An overall increase in the level of TCRBV 2 transcripts was detected in ALS specimens when compared to controls. This result was independent of the HLA genotype of the individual. Furthermore, enrichment of TCRBV2-positive T cells could be demonstrated in cerebrospinal fluid derived from patients with ALS, using PCR analysis and a T cell stimulation assay with toxic shock syndrome toxin-1 (TSST-1), a Vbeta2-specific superantigen. Our results suggest that an immunological process involving the specific expansion of Vbeta2 TCR-positive T-cells may be important in the pathogenesis of ALS.