BACKGROUND C-X-C chemokine receptor type 5(CXCR5)+CD8+T cells represent a unique immune subset with dual roles,functioning as cytotoxic cells in persistent viral infections while promoting B cell responses.Despite the...BACKGROUND C-X-C chemokine receptor type 5(CXCR5)+CD8+T cells represent a unique immune subset with dual roles,functioning as cytotoxic cells in persistent viral infections while promoting B cell responses.Despite their importance,the specific role of CXCR5+CD8+T cells in chronic hepatitis B(CHB),particularly during interferon-alpha(IFN-α)treatment,is not fully understood.This study aims to elucidate the relationship between CXCR5+CD8+T cells and sustained serologic response(SR)in patients undergoing 48 weeks of pegylated IFN-α(peg-IFN-α)treatment for CHB.AIM To elucidate the relationship between CXCR5+CD8+T cells and sustained SR in patients undergoing 48 weeks of peg-IFN-αtreatment for CHB.METHODS This study enrolled 60 patients with hepatitis Be antigen(HBeAg)-positive CHB undergoing 48 weeks of peg-IFN-αtreatment.Participants were assessed for eligibility based on criteria such as persistent HBsAg-positive status for at least six months,HBeAb-negative,hepatitis B virus DNA levels exceeding 2×104 copies/mL,and alanine aminotransferase(ALT)levels between 2 and 10 times the upper limit of normal.Blood samples were collected at baseline and at weeks 12,24,48,and a 24-week treatment-free follow-up(week 72)to measure serum interleukin(IL)-21 concentration via ELISA and to analyze CXCR5 and programmed death-ligand 1(PD-L1)expression on CD8+T cells by flow cytometry,CXCR5 is a chemokine receptor that directs immune cells to specific tissues,while PD-L1 is a protein that regulates immune responses by inhibiting T cell activity.RESULTS Patients with CHB exhibited significantly lower levels of circulating CXCR5+CD8+T cells compared to healthy controls(P<0.01).Notably,CXCR5+CD8+T cells were prominently expressed in patients who achieved sustained SR compared to non-SR(NSR).A significant correlation was observed between CXCR5 and PD-L1 expression(r=-0.189,P=0.002).However,there was no significant correlation between serum IL-21 levels and CXCR5+CD8+lymphocytes(r=-0.03,P=0.625)or serum ALT levels(r=0.026,P=0.678).CONCLUSION The enhanced expression of CXCR5+CD8+T cells in patients achieving HBeAg seroconversion during IFN-αtreatment suggests that these cells play a crucial role in antiviral immune responses against hepatitis B.This study highlights the potential of CXCR5+CD8+T cells as immune regulators in CHB,which may inform future therapeutic strategies to optimize antiviral treatments.展开更多
Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemok...Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemokine superfamily receptors were aligned by computer and the least homologous domain of the extracellular loops was located. In this paper, the first extracellular domain (114 nucleotides) was defined and amplified by PCR, and a expression vector of the recombinant GST fusion protein,pGEXIN/NR5, was constructed. After the inserted sequence was confirmed correct, the transformation and expression of fusion protein was performed in E. coli and the fusion protein was purified. Then 2 Newzealand rabbits were immunized. The anti-CCR5 NH2-terminal antibody F(ab’ )2 was made by protein A affinity chromatography, pepsin digestion and sepharose-12 column chromatography. Results: Reduced and unreduced SDS-PAGE and FAX analysis demonstrated that this F(ab’ )2 had a high specificity to combine with CCR5. Conclusion: A simple and quick method to prepare specific antibody F(ab’ )2 of certain functional domain is showed in this paper, by which we can get an improtant experiment material for studying gene expression, and it will provide a good idea and a technique to study other high similar superfamily members.展开更多
基金Supported by Changsha Science and Technology Program,No.kq2022397Natural Science Foundation of Hunan Province(Departmental Joint Fund),No.2023JJ60440+2 种基金Research Program of Health Commission of Hunan Province,No.202303088786Clinical Medical Research Center for Viral Hepatitis of Hunan Province,No.2023SK4009the Scientific Research Program of FuRong Laboratory,No.2023SK2108.
文摘BACKGROUND C-X-C chemokine receptor type 5(CXCR5)+CD8+T cells represent a unique immune subset with dual roles,functioning as cytotoxic cells in persistent viral infections while promoting B cell responses.Despite their importance,the specific role of CXCR5+CD8+T cells in chronic hepatitis B(CHB),particularly during interferon-alpha(IFN-α)treatment,is not fully understood.This study aims to elucidate the relationship between CXCR5+CD8+T cells and sustained serologic response(SR)in patients undergoing 48 weeks of pegylated IFN-α(peg-IFN-α)treatment for CHB.AIM To elucidate the relationship between CXCR5+CD8+T cells and sustained SR in patients undergoing 48 weeks of peg-IFN-αtreatment for CHB.METHODS This study enrolled 60 patients with hepatitis Be antigen(HBeAg)-positive CHB undergoing 48 weeks of peg-IFN-αtreatment.Participants were assessed for eligibility based on criteria such as persistent HBsAg-positive status for at least six months,HBeAb-negative,hepatitis B virus DNA levels exceeding 2×104 copies/mL,and alanine aminotransferase(ALT)levels between 2 and 10 times the upper limit of normal.Blood samples were collected at baseline and at weeks 12,24,48,and a 24-week treatment-free follow-up(week 72)to measure serum interleukin(IL)-21 concentration via ELISA and to analyze CXCR5 and programmed death-ligand 1(PD-L1)expression on CD8+T cells by flow cytometry,CXCR5 is a chemokine receptor that directs immune cells to specific tissues,while PD-L1 is a protein that regulates immune responses by inhibiting T cell activity.RESULTS Patients with CHB exhibited significantly lower levels of circulating CXCR5+CD8+T cells compared to healthy controls(P<0.01).Notably,CXCR5+CD8+T cells were prominently expressed in patients who achieved sustained SR compared to non-SR(NSR).A significant correlation was observed between CXCR5 and PD-L1 expression(r=-0.189,P=0.002).However,there was no significant correlation between serum IL-21 levels and CXCR5+CD8+lymphocytes(r=-0.03,P=0.625)or serum ALT levels(r=0.026,P=0.678).CONCLUSION The enhanced expression of CXCR5+CD8+T cells in patients achieving HBeAg seroconversion during IFN-αtreatment suggests that these cells play a crucial role in antiviral immune responses against hepatitis B.This study highlights the potential of CXCR5+CD8+T cells as immune regulators in CHB,which may inform future therapeutic strategies to optimize antiviral treatments.
文摘Objective: To study the function of the first extracellular membrane loop of the NH2-terminal of chemokine receptor 5(CCRS), and produce its specific antibody F(ab’ )2. Methods: Some amino acid sequences of β chemokine superfamily receptors were aligned by computer and the least homologous domain of the extracellular loops was located. In this paper, the first extracellular domain (114 nucleotides) was defined and amplified by PCR, and a expression vector of the recombinant GST fusion protein,pGEXIN/NR5, was constructed. After the inserted sequence was confirmed correct, the transformation and expression of fusion protein was performed in E. coli and the fusion protein was purified. Then 2 Newzealand rabbits were immunized. The anti-CCR5 NH2-terminal antibody F(ab’ )2 was made by protein A affinity chromatography, pepsin digestion and sepharose-12 column chromatography. Results: Reduced and unreduced SDS-PAGE and FAX analysis demonstrated that this F(ab’ )2 had a high specificity to combine with CCR5. Conclusion: A simple and quick method to prepare specific antibody F(ab’ )2 of certain functional domain is showed in this paper, by which we can get an improtant experiment material for studying gene expression, and it will provide a good idea and a technique to study other high similar superfamily members.
