|
|
Evaluation on the immunogenicity of the antigen epitopes concentrated areas of Rv2941 protein from Mycobacterium tuberculosis |
FAN Xue-ting1, LUAN Xiu-li1, ZHAO Xiu-qin1, LI Ma-chao1, WAN Kang-lin1, LU Xuan-cheng2, LI Xiao-yan2, LIU Hai-can1 |
1. State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; 2. Laboratory Animal Center, Chinese Center for Disease Control and Prevention, Beijing 102206, China |
|
|
Abstract To analyze the immunogenicity of the antigen epitopes concentrated areas of Rv2941 protein from Mycobacterium tuberculosis, and investigate its potential candidate antigen for tuberculosis vaccine. T cell epitopes of Rv2941 antigen was analyzed by Immune Epitope Database (IEDB). The antigen epitopes concentrated areas (649-1 017 bp, named Rv2941p) of Rv2941 gene was inserted into pET32a vector, then expressed in Escherichia coli (E.coli) (DE3) BL21 cells. Prokaryotic expression showed that this protein was mainly in the form of soluble form. BALB/c mice were immunized subcutaneously in triplicate over a 10-day interval. One week after the last immunization, the samples were collected to assess the immunogenicity. Rv2941p elicited significantly higher antigen-specific immunoglobulin G (IgG) than control groups (P<0.001), indicating that Rv2941p enhanced antibody response. To evaluate the cell mediated immune response, the cytokines (e.g., IL-4, IL-2, IL-6 and IFN-γ) were detected by ELISA. These results showed that Rv2941p significantly improved the level of IFN-γ and IL-6 compared with Ag85B (P<0.001). Meanwhile, the results of IgG subclass shown that Rv2941p could improve the ratio of IgG2a/IgG1. And the percentage of splenic CD4+T and CD8+T cells were measured by flow cytometry. In addition, the intracellular cytokines (e.g., IFN-γ, TNF-α and IL-4) were detected by flow cytometry. We found that Rv2941p enhanced intracellular expression of IFN-γ and TNF-α cytokines, and promoted CD4+ T cell and CD8+T cell proliferation. Further this study provided evidence that the T cell epitopes of Rv2941 promoted stronger cell-mediated immune response, and skewed to Th-1 type cellular immune response. In conclusion, the T cell epitope of Rv2941 could promote humoral and cell-mediated immune responses, which supports it as a promising antigen for TB vaccines.
|
Received: 07 March 2022
|
|
Corresponding Authors:
Liu Hai-can, Email:liuhaican@icdc.cn; Li Xiao-yan, Email: lixy@chinacdc.cn
|
|
|
|
[1] Karbalaei Zadeh Babaki M, Soleimanpour S, Rezaee SA. Antigen 85 complex as a powerful Mycobacterium tuberculosis immunogene: Biology, immune-pathogenicity, applications in diagnosis, and vaccine design[J]. Microb Pathog, 2017, 112: 20-29. DOI:10.1016/j.micpath.2017.08.040 [2] Steingart KR, Ng V, Henry M, et al.Sputum processing methods to improve the sensitivity of smear microscopy for tuberculosis: a systematic review[J]. Lancet Infect Dis, 2006, 6(10): 664-674. DOI:10.1016/S1473-3099(06) 70602-8 [3] Houben RM, Dodd PJ.The global burden of latent tuberculosis infection: a re-estimation using mathematical modelling[J]. PLoS Med, 2016, 13(10): e1002152. DOI:10.1371/journal.pmed.1002152 [4] Fine PE.Variation in protection by BCG: implications of and for heterologous immunity[J]. Lancet, 1995, 346(8986):1339-1345. DOI:10.1016/s0140-6736(95)92348-9 [5] Lin MY, Geluk A, Smith SG, et al.Lack of immune responses to Mycobacterium tuberculosis DosR regulon proteins following Mycobacterium bovis BCG vaccination[J]. Infect Immun, 2007, 75(7): 3523-3530. DOI:10.1128/IAI.01999-06 [6] Aronson NE, Santosham M, Comstock GW, et al.Long-term efficacy of BCG vaccine in American Indians and Alaska Natives: a 60-year follow-up study[J]. JAMA, 2004, 291(17): 2086-2091. DOI:10.1001/jama.291.17.2086 [7] Levillain F, Kim H, Woong Kwon K, et al.Preclinical assessment of a new live attenuated Mycobacterium tuberculosis Beijing-based vaccine for tuberculosis[J]. Vaccine, 2020, 38(6): 1416-1423. DOI:10.1016/j.vaccine. 2019.11.085 [8] Nguipdop-Djomo P, Heldal E, Rodrigues LC, et al.Duration of BCG protection against tuberculosis and change in effectiveness with time since vaccination in Norway: a retrospective population-based cohort study[J]. Lancet Infect Dis, 2016, 16(2): 219-226. DOI:10.1016/S1473-3099(15)00400-4 [9] Goyal A, Yousuf M, Rajakumara E, et al.Crystallization and preliminary X-ray crystallographic studies of the N-terminal domain of FadD28, a fatty-acyl AMP ligase from Mycobacterium tuberculosis[J]. Acta Crystallogr Sect F Struct Biol Cryst Commun, 2006, 62(Pt4):350-352. DOI:10.1107/S1744309106005938 [10] Madan R, Pandit K, Bhati L, et al.Mining the Mycobacterium tuberculosis proteome for identification of potential T-cell epitope based vaccine candidates[J]. Microb Pathog, 2021, 157: 104996. DOI:10.1016/j.micpath. 2021.104996 [11] Sirakova TD, Fitzmaurice AM, Kolattukudy P.Regulation of expression of mas and fadD28, two genes involved in production of dimycocerosyl phthiocerol, a virulence factor of Mycobacterium tuberculosis[J]. J Bacteriol, 2002, 184(24): 6796-6802. DOI:10.1128/JB.184.24.6796-6802.2002 [12] Lindestam Arlehamn CS, Lewinsohn D, Sette A, et al.Antigens for CD4 and CD8 T cells in tuberculosis[J]. Cold Spring Harb Perspect Med, 2014, 4(7): a018465. DOI:10.1101/cshperspect.a018465 [13] Orr MT, Duthie MS, Windish HP, et al.MyD88 and TRIF synergistic interaction is required for TH1-cell polarization with a synthetic TLR4 agonist adjuvant[J]. Eur J Immunol, 2013, 43(9): 2398-2408. DOI:10.1002/eji.201243124 [14] Leal IS, Smedegard B, Andersen P, et al.Interleukin-6 and interleukin-12 participate in induction of a type 1 protective T-cell response during vaccination with a tuberculosis subunit vaccine[J]. Infect Immun, 1999, 67(11): 5747-5754. DOI: 10.1128/IAI.67.11.5747-5754.1999 [15] Sia JK, Rengarajan J.Immunology of Mycobacterium tuberculosis infections[J]. Microbiol Spectr, 2019, 7(4):GPP3-0022-2018. DOI:10.1128/microbiolspec.GPP3-0022-2018 [16] Portevin D, Gagneux S, Comas I, et al.Human macrophage responses to clinical isolates from the Mycobacterium tuberculosis complex discriminate between ancient and modern lineages[J]. PLoS Pathog, 2011. 7(3): e1001307. DOI:10.1371/journal.ppat.1001307 [17] Li J, Jin C, Wu C, et al.PD-1 modulating Mycobacterium tuberculosis-specific polarized effector memory T cells response in tuberculosis pleurisy[J]. J Leukoc Biol, 2019, 106(3):733-747. DOI: 10.1002/JLB.MA1118-450RR [18] Slight SR, Rangel-Moreno J, Gopal R, et al.CXCR5+ T helper cells mediate protective immunity against tuberculosis[J]. J Clin Invest, 2013, 123(2): 712-726. DOI:10.1172/JCI65728 |
[1] |
BAI Lu, NING Huan-huan, KANG Jian, LIANG Xuan, XIE Yan-ling, PENG Yu-jun, ZHANG Jing-yao, LU Yan-zhi, BAI Yin-lan. Immune responses induced by mucosal immunization with Mycobacterium tuberculosis EsxV subunit vaccine[J]. Chinese Journal of Zoonoses, 2022, 38(5): 379-386. |
|
|
|
|