ABSTRACT
Objective:To study the protective effects of bicistronic DNA vaccines carrying herpes simplex virus type 2 glycoprotein D (HSV-2 gD) and adjuvant CCL28 sequences that were connected by internal ribosome entry site (IRES) sequence in mouse model.Methods:The recombinant DNA vaccines, pgD-IRES-CCL28 and pCCL28-IRES-gD, encoding HSV-2 gD and adjuvant CCL28 were constructed with IRES sequence. After verified by sequencing, they were intramuscularly injected twice into BALB/c mice. Serum samples and vaginal lavage fluids were collected regularly. Splenocytes, mesenteric lymph node cells and rectal mucosa tissues were separated and collected. The titers of antigen-specific antibodies in immunized mice were analyzed with end-point ELISA. In vitro neutralization assay was used to measure neutralizing antibody titers in serum and vaginal lavage fluid after vaccination and virus challenge. CCL28-responsive immune cells in splenocytes, mesenteric lymph node cells and rectal tissues were detected by chemotaxis experiment and immunohistochemical staining. The protective effects of the bicistronic DNA vaccines were evaluated by fluorescent quantitative PCR, weighing and disease severity assessment. Humoral and cellular immune responses induced by the bicistronic DNA vaccines and their efficacy in immunoprotection were analyzed by comparing with pgD+ pCCL28 group. Results:IgG titers in serum samples and IgA antibody titers in vaginal lavage fluids of mice immunized with pCCL28-IRES-gD were similar to those in pgD+ pCCL28 group. The neutralizing ability of antibodies, the number of rectal mucosal IgA+ plasma cells and CCL28-responsive immune cells in mucosal tissues were increased in pCCL28-IRES-gD group. Serum neutralizing antibodies were not produced immediately in the mice challenged with HSV-2, but no weight loss, disease symptoms or death was observed. However, pgD+ pcDNA3.1 and pgD-IRES-CCL28 were ineffective against HSV-2 infection in mice.Conclusions:The recombinant bicistronic DNA vaccine of pCCL28-IRES-gD could induce stronger mucosal immune response in mice and provide better protective effects.
ABSTRACT
Objective To investigate the expression of chemokine CCL27,CCL28 and their receptor CCR10 in mouse periph-eral blood in the acute phase of epilepsy.Methods The peripheral blood of acute epileptic mice at different time points(10 min,30 min,1 h,2 h)was collected,real-time PCR was used to detect the mRNA expression level of CCL27 and CCL28.The heparin anti-coagulation peripheral blood at the same time points(10 min,30 min,1 h,2 h)of normal and acute phase of epileptic mice were col-lected and flow cytometry was used to investigate the expression of CCR10 in peripheral blood lymphocyte.Results The mRNA expression level of CCL27,CCL28 in peripheral blood and the expression of CCR10 in lymphocytes were found significantly in-creased at 2 h in epileptic mice than those of normal(P <0.01).Conclusion The immune function disorder occured in peripheral blood in early epilepsic pathological process and might be associated with the subsequent inflammatory reaction and neuron apoptosis.
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Background and purpose:Chemokines play an important role in breast cancer occurrence and development. However, little about the function of CCL28 in breast cancer is reported. This study was designed to observe and study the expression of chemokine CCL28 in breast cancer, and analyze their association with pathological features of breast cancer.Methods:Expressions of chemokine CCL28 in 150 breast cancer patients were determined by IHC(immunohistochemistry) technique. The corresponding normal breast tissues in paraneoplastic were also detected. The level of CCL28 expression in 150 breast cancer was analyzed whether it was associated with age, cTNM stage, tumor diameter, axillary lymph node status, ER status, PR status and HER-2 status. Results:①CCL28 was highly expressed in both breast cancer and normal breast tissues in paraneoplastic. The positive expression rate of CCL28 in breast cancer was 54.6%and the positive expression rate of CCL28 in normal paraneoplastic breast tissues was 9.3%. Expression of CCL28 in breast cancer was signiifcantly higher than that in normal paraneoplastic breast tissues, there was statistically significance between the two groups(P0.05). Conclusion:The level of CCL28 expression showed signiifcantly difference between breast cancer and paraneoplastic normal breast tissues. In conclusion, CCL28 may be correlated with breast cancer carcinogenesis and evolvement. The level of CCL28 expression was not signiifcantly correlated with lymph node metastasis. If CCL28 may be a factor to predict lymph node metastasis in breast cancer is worth further studying.