Pathogens
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2021
·Vol. 11
·期 1
·7
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IF 3.492
Fuchun Zhang,Lingling Sun,Mark K. Lafferty,Joseph B. Margolick,Alfredo Garzino-Demo
CD4+CCR6+T cells are highly susceptible to HIV infection, and a high cytokine producing CCR6+T cell subset is selectively lost during HIV infection. The CCR6 chemokine MIP-3α (CCL20) is produced at sites of infection in SIV animal models. Recently, we have shown that MIP-3α inhibits HIV replication. This inhibition of HIV infection is mediated by CCR6 signaling and eventuates in increased APOBEC3G expression. Since there are few existing reports on the role of MIP-3α in health or disease, we studied its production by PBMCs from HIV-seronegative and HIV+ subjects. We evaluated the ability of PBMCs to produce MIP-3α in response to antigen stimulation using cells obtained from two groups: one composed of HIV-seronegative subjects (n = 16) and the other composed of HIV+ subjects (n = 58), some asymptomatic and some with clinically defined AIDS. Antigens included fragment C of the tetanus toxin,Candida albicans, whole-inactivated HIV, and HIV p24. MIP-3α was detected by ELISA in tissue culture supernatants of antigen-stimulated PBMCs. MIP-3α production by antigen-stimulated PBMCs was readily measured for HIV-negative subjects and for HIV-seropositive asymptomatic subjects, but not for patients with AIDS. These results suggest that subversion of the MIP-3α-CCR6 axis by HIV during the course of infection contributes to the loss of immune function that eventually leads to AIDS.Keywords:CCR6;MIP-3α;immune response;antigen stimulation;Th17
研究领域:
免疫学
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病毒学
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DOI
PubMed
高影响因子
Drug Design Development and Therapy
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2026
·
IF 6.1
Na Yang,Yuqing Zhao,Shiyu Liu,Yifan Zhou,Xinyue Ji,Tianqi Zhang,Runbin Sun,Huaijun Zhu,Yao Du,Min Wang
Background Mycophenolic acid (MPA), administered as mycophenolate mofetil (MMF) or enteric-coated mycophenolate sodium (EC-MPS), is the first-line immunosuppressant for kidney transplant patients. Traditional plasma-based therapeutic drug monitoring (TDM) for MPA fails to accurately reflect intracellular drug exposure at the pharmacological action site.Purpose This study aimed to establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of MPA and its glucuronide metabolite, MPAG, in peripheral blood mononuclear cells (PBMCs) to support cellular pharmacokinetic (PK) assessment.Methods Chromatographic separation was performed on a BEH C18 column using methanol-water containing 0.1% formic acid and 5 mmol/L ammonium acetate. A total of 139 PBMC samples from 40 kidney transplant recipients (n=19 receiving EC-MPS; n=21 receiving MMF) were analyzed to evaluate the plasma-PBMC PK correlations and gender-related differences.Results The method exhibited excellent linearity over the concentration range of 0.2–500 ng/mL (R2 > 0.998), along with acceptable precision, accuracy and matrix effect. The analytes remained stable under various experimental conditions. Significant inter-individual variability in intracellular MPA and MPAG concentrations was observed among patients. Spearman correlation analysis demonstrated a weak-to-moderate positive correlation between plasma and intracellular PBMC concentrations of MPA (EC-MPS, Rs=0.396; MMF, Rs=0.480). Additionally, the concentration-time profiles of MPA and MPAG in PBMCs of kidney transplant recipients after administration of EC-MPS or MMF exhibited different characteristics. No significant gender-based differences in plasma and PBMCs were observed.Conclusion We established and validated an LC-MS/MS method for simultaneous quantification of intracellular MPA and MPAG. The observed weak-to-moderate correlation between plasma and PBMC exposure highlights the inherent limitations of conventional plasma-based TDM for MPA. Collectively, our study provides a feasible approach for monitoring intracellular drug levels and facilitates the optimization of individualized immunosuppressive dosing regimens for renal transplant recipients.
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DOI
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2024
·
IF 4.8
Shengsheng Mei,Shanshan Peng,Eu Gene Vong,Jinbiao Zhan
Background Clinical data indicate that a substantial portion of cancer patients, though eligible for immune checkpoint inhibitor (ICI) therapy, cannot fully benefit from ICI monotherapy due to the poor immunogenicity of tumors and lack of tumor-infiltrating lymphocytes within the ‘cold’ tumor microenvironment (TME). In addition to poor antibody penetrance into the TME, systemic delivery of ICIs is associated with immune-related adverse events (irAEs) among recipients, some of which are life-threatening. Oncolytic virotherapy is a potentially viable approach to improving the efficacy of ICI therapy because of their ability to selectively replicate and lyse tumor cells, release tumor-associated antigens (TAAs), induce inflammatory response and promote lymphocyte infiltration in tumors. Methods A recombinant oncolytic adenoviruses (OAd), denoted ZD55-aPD-L1 scFv, which carried the expression cassette for anti-PD-L1 scFv was constructed by molecular cloning. Western blot and ELISA assay were performed to detect aPD-L1 scFv expression. Flow cytometry were used to analyse PD-L1 expression and count tumor cells. Co-culture assay of human peripheral blood mononuclear cells (PBMCs) with tumor cells in vitro and triple-negative breast cancer (TNBC) MDA-MB-231 tumor-bearing model in vivo were evaluated the antitumor effects of recombinant oncolytic adenoviruses ZD55-aPD-L1 scFv. Results We found that cells infected with recombinant oncolytic adenovirus ZD55-aPD-L1 scFv can effectively express aPD-L1 scFv, which function similarly to its full-length anti-PD-L1 antibody. PBMCs have inherently very limited killing effect on tumor cells even with administration of anti-PD-L1 antibody as observed from our in vitro co-cultures. Treatment consisting of ZD55 alone or ZD55 combined with anti-PD-L1 antibody yielded mediocre antitumor efficacy in subsequent in vitro and in vivo investigations, but were all substantially surpassed by the synergistic antitumor effects observed with ZD55-aPD-L1 scFv treatment. We show that the concomitant direct oncolysis by the recombinant OAd and localized autocrine/paracrine interception of PD-1:PD-L1 checkpoint interaction mediated by ZD55-aPD-L1 scFv-infected cells is exceedingly superior to co-administration of ZD55 and anti-PD-L1 antibody in the human TNBC mice model. Conclusions Our results provided evidence for the development of novel strategies, in this case an anti-PD-L1 scFv-armed OAd, to bolster immune responses to ‘cold’ tumors and to improve therapeutic responsiveness to ICIs.
研究领域:
肿瘤学
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免疫疗法
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分子生物学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
EXPERIMENTAL PARASITOLOGY
·
2023
·
IF 2.1
Muhammad Ehsan,Rui-Si Hu,Meng Wang,Jun-Ling Hou,Muhammad Rashid,Muhammad Irfan Malik
Legumains belonging to C_13 peptidase family of proteins, and are ubiquitously disseminated among all vertebrate and invertebrate organisms, and have been implicated in innumerable biological and cellular functionality. Herein, we characterized and evaluated immunoregulatory characteristics of Legumain-1 from Fasciola gigantica (Fg-LGMN-1) during its interaction with host immune cells . The isopropyl-ß-d-thiogalactopyranoside (IPTG) stimulated RFg-LGMN-1 protein was positively detected by rat serum containing anti-RFg-LGMN-1 polyclonal antibodies . Furthermore, the uptake of RFg-LGMN-1 by goat monocytes was successfully confirmed using Immunofluorescence Assay (IFA). The immunohistochemical analysis revealed the native localization of LGMN-1 protein on the periphery and internal structures such as suckers, pharynx, and genital pore of the adult parasite, thereby validating its presence in excretory-secretory (ES) products of F. gigantica . The RFg-LGMN-1 co-incubated with concanavalin-A (Con-A) stimulated the increase of interleukin 2 (IL-2), IL-10, and IL-17 in monocytes derived from peripheral blood mononuclear cells (PBMCs) in the concentration-dependent manner. However, the IL-4 cytokine in response to the RFg-LGMN-1 protein declined. These results illuminated the role of LGMN-1 during the parasite-host interface. Our findings elaborated additional evidence that Legumain protein play a role in the manipulating host immune responses during parasite infections. However, further evaluation of RFg-LGMN-1 protein in context of its immunomodulatory roles should be conducted to enhance our understandings of the mechanisms employed by F. gigantica to evade host immune responses.
研究领域:
免疫学
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寄生虫学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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