Preparation and Application of Fluorescent Monoclonal Antibodies Recognizing Goat CD4CD25 Regulatory T Cells

Overview

Journal Appl Microbiol Biotechnol

Specialties Biomedical Engineering
Biotechnology
Microbiology

Date 2024 May 8

PMID 38717623

Authors

Yunpeng Wang

Haoyue Yang

Jiajin Hu

Yuecai Jiang

Wentao Ma

Shikong Gao

Dekun Chen

Affiliations

Soon will be listed here.

Abstract

Regulatory T cells (Tregs) are a subset of T cells participating in a variety of diseases including mycoplasmal pneumonia, contagious ecthyma, and so on. The role of Tregs in goat contagious ecthyma is not completely understood due to the lack of species-specific antibodies. Here, we developed a combination of CD4 and CD25 fluorescence monoclonal antibodies (mAb) to recognize goat Tregs and assessed its utility in flow cytometry, immunofluorescence staining. Using immunofluorescence staining, we found that the frequency of Treg cells was positively correlated with the viral load during orf virus infection. These antibodies could serve as important tools to monitor Tregs during orf virus infection in goats. KEY POINTS: • A combination of fluorescent mAbs (C11 and D12) was prepared for the detection of goat Tregs. • C11 and D12 are effective in flow cytometry, immunofluorescence staining, and C11 has excellent species specificity. • The frequency of Treg cells was positively correlated with the viral load during orf virus infection.

References

Lawan Z, Bala J, Bukar A, Balakrishnan K, Mangga H, Abdullah F . Contagious ecthyma: how serious is the disease worldwide?. Anim Health Res Rev. 2021; 22(1):40-55. DOI: 10.1017/S1466252320000018. View

Farooq A, Shukla D . Herpes simplex epithelial and stromal keratitis: an epidemiologic update. Surv Ophthalmol. 2012; 57(5):448-62. PMC: 3652623. DOI: 10.1016/j.survophthal.2012.01.005. View

Cheung J, Zahorowska B, Suranyi M, Wong J, Diep J, Spicer S . CD4CD25 T regulatory cells in renal transplantation. Front Immunol. 2022; 13:1017683. PMC: 9681496. DOI: 10.3389/fimmu.2022.1017683. View

Ghazaleh R, Al-Sawalhe M, Abu Odeh I, El Ibrahim J, Al-Turman B, Makhamreh J . Host range, severity and trans boundary transmission of Orf virus (ORFV). Infect Genet Evol. 2023; 112:105448. DOI: 10.1016/j.meegid.2023.105448. View

Liu Y, Beyer A, Aebersold R . On the Dependency of Cellular Protein Levels on mRNA Abundance. Cell. 2016; 165(3):535-50. DOI: 10.1016/j.cell.2016.03.014. View

Bodi I, H-Minko K, Prodan Z, Nagy N, Olah I . [Structure of the thymus at the beginning of the 21th century]. Orv Hetil. 2019; 160(5):163-171. DOI: 10.1556/650.2019.31224. View

Gangaplara A, Martens C, Dahlstrom E, Metidji A, Gokhale A, Glass D . Type I interferon signaling attenuates regulatory T cell function in viral infection and in the tumor microenvironment. PLoS Pathog. 2018; 14(4):e1006985. PMC: 5929570. DOI: 10.1371/journal.ppat.1006985. View

Jansen K, Wirz O, Veen W, Tan G, Mirer D, Sokolowska M . Loss of regulatory capacity in Treg cells following rhinovirus infection. J Allergy Clin Immunol. 2021; 148(4):1016-1029.e16. DOI: 10.1016/j.jaci.2021.05.045. View

Levings M, Sangregorio R, Sartirana C, Moschin A, Battaglia M, Orban P . Human CD25+CD4+ T suppressor cell clones produce transforming growth factor beta, but not interleukin 10, and are distinct from type 1 T regulatory cells. J Exp Med. 2002; 196(10):1335-46. PMC: 2193983. DOI: 10.1084/jem.20021139. View

10.

Chen M, Pittet M, Gorelik L, Flavell R, Weissleder R, von Boehmer H . Regulatory T cells suppress tumor-specific CD8 T cell cytotoxicity through TGF-beta signals in vivo. Proc Natl Acad Sci U S A. 2004; 102(2):419-24. PMC: 544311. DOI: 10.1073/pnas.0408197102. View

11.

Sebastianelli M, Pavone S, Forte C, Mezzasoma S, Bordoni M, Onofri A . Preliminar histological study of the thymus in regularly slaughtered Chianina beef cattle to improve methods for the detection of illicit treatments. Vet Ital. 2020; 56(2):137-140. DOI: 10.12834/VetIt.1624.8712.1. View

12.

Oldenhove G, Bouladoux N, Wohlfert E, Hall J, Chou D, Dos Santos L . Decrease of Foxp3+ Treg cell number and acquisition of effector cell phenotype during lethal infection. Immunity. 2009; 31(5):772-86. PMC: 2814877. DOI: 10.1016/j.immuni.2009.10.001. View

13.

Rubtsov Y, Rasmussen J, Chi E, Fontenot J, Castelli L, Ye X . Regulatory T cell-derived interleukin-10 limits inflammation at environmental interfaces. Immunity. 2008; 28(4):546-58. DOI: 10.1016/j.immuni.2008.02.017. View

14.

Li Y, Wang Q, Bergmann S, Zeng W, Wang Y, Ren Y . Preparation of monoclonal antibodies against KHV and establishment of an antigen sandwich ELISA for KHV detection. Microb Pathog. 2018; 128:36-40. DOI: 10.1016/j.micpath.2018.12.034. View

15.

Haig D, Mercer A . Ovine diseases. Orf. Vet Res. 1998; 29(3-4):311-26. View

16.

Gao Y, Sang F, Meng D, Wang Y, Ma W, Chen D . Preparation of a novel monoclonal antibody against caprine interleukin-17A and its applications in immunofluorescence and immunohistochemistry assays. BMC Biotechnol. 2019; 19(1):47. PMC: 6637523. DOI: 10.1186/s12896-019-0543-5. View

17.

Wagner B, Hillegas J, Antczak D . A monoclonal antibody to equine interleukin-4. Vet Immunol Immunopathol. 2006; 110(3-4):363-7. DOI: 10.1016/j.vetimm.2006.01.001. View

18.

Yu W, Geng S, Suo Y, Wei X, Cai Q, Wu B . Critical Role of Regulatory T Cells in the Latency and Stress-Induced Reactivation of HSV-1. Cell Rep. 2018; 25(9):2379-2389.e3. DOI: 10.1016/j.celrep.2018.10.105. View

19.

Hang L, Kumar S, Blum A, Urban Jr J, Fantini M, Weinstock J . Infection Decreases Smad7 Expression in Intestinal CD4 T Cells, Which Allows TGF-β to Induce IL-10-Producing Regulatory T Cells That Block Colitis. J Immunol. 2019; 202(8):2473-2481. PMC: 7890536. DOI: 10.4049/jimmunol.1801392. View

20.

Qiu R, Zhou L, Ma Y, Zhou L, Liang T, Shi L . Regulatory T Cell Plasticity and Stability and Autoimmune Diseases. Clin Rev Allergy Immunol. 2018; 58(1):52-70. DOI: 10.1007/s12016-018-8721-0. View