Polyclonal Antibody-based Immunohistochemical Detection of Intraleukocytic Parasites in Roan and Sable Antelopes

Overview

Journal J Vet Diagn Invest

Specialty Veterinary Medicine

Date 2021 Aug 2

PMID 34333997

Authors

Sarah J Clift

Bernat Marti-Garcia

Rephima M Phaswane

Emily P Mitchell

Antoinette I Josemans

Ilse Vorster

Katja N Koeppel

Jeanni Fehrsen

Affiliations

Soon will be listed here.

Abstract

parasites commonly infect African wild artiodactyls. In rare roan () and sable () antelopes, sp. (sable)-associated calf mortalities constrain breeding programs. The pathogenicity of most leukocyte-transforming spp. originates in their invasion of and multiplication in various mononuclear leukocytes, the transformation of both infected and uninfected leukocytes, and their infiltration of multiple organs. Understanding the pathogenesis of theileriosis can be improved by the use of immunohistochemistry (IHC) to identify the localization of the parasites in tissue sections. Our aim was to develop a reproducible IHC assay to detect leukocyte-associated parasites in formalin-fixed, paraffin-embedded roan and sable tissues. Polyclonal antibodies were purified from the sera of 5 roans from an area endemic for sp. (sable) and tested for IHC reactivity in 55 infected and 39 control roan and sable antelopes, and for antigen and species cross-reactivity in an additional 58 cases. The 3 strongest antibodies consistently detected intraleukocytic theilerial antigens in known positive cases in roan and sable antelopes, and also detected other spp. in non-hippotraginid wild artiodactyl tissues. The antibodies did not cross-react with other apicomplexan protozoa, with the exception of . Given that PCR on its own cannot determine the significance of theilerial infection in wild ruminants, IHC is a useful laboratory test with which to confirm the diagnosis in these species.

References

Ramos-Vara J, Kiupel M, Baszler T, Bliven L, Brodersen B, Chelack B . Suggested guidelines for immunohistochemical techniques in veterinary diagnostic laboratories. J Vet Diagn Invest. 2008; 20(4):393-413. DOI: 10.1177/104063870802000401. View

Branco S, Orvalho J, Leitao A, Pereira I, Malta M, Mariano I . Fatal cases of Theileria annulata infection in calves in Portugal associated with neoplastic-like lymphoid cell proliferation. J Vet Sci. 2010; 11(1):27-34. PMC: 2833427. DOI: 10.4142/jvs.2010.11.1.27. View

Holden P, Horton W . Crude subcellular fractionation of cultured mammalian cell lines. BMC Res Notes. 2009; 2:243. PMC: 2802353. DOI: 10.1186/1756-0500-2-243. View

Forsyth L, Minns F, Kirvar E, Adamson R, Hall F, McOrist S . Tissue damage in cattle infected with Theileria annulata accompanied by metastasis of cytokine-producing, schizont-infected mononuclear phagocytes. J Comp Pathol. 1999; 120(1):39-57. DOI: 10.1053/jcpa.1998.0256. View

Buchwalow I, Samoilova V, Boecker W, Tiemann M . Non-specific binding of antibodies in immunohistochemistry: fallacies and facts. Sci Rep. 2012; 1:28. PMC: 3216515. DOI: 10.1038/srep00028. View

Gharbi M, Darghouth M, Elati K, Al-Hosary A, Ayadi O, Salih D . Current status of tropical theileriosis in Northern Africa: A review of recent epidemiological investigations and implications for control. Transbound Emerg Dis. 2020; 67 Suppl 1:8-25. DOI: 10.1111/tbed.13312. View

Clift S, Collins N, Oosthuizen M, Steyl J, Lawrence J, Mitchell E . The Pathology of Pathogenic Theileriosis in African Wild Artiodactyls. Vet Pathol. 2019; 57(1):24-48. DOI: 10.1177/0300985819879443. View

Emery D . Kinetics of infection with Theileria parva (East Coast fever) in the central lymph of cattle. Vet Parasitol. 1981; 9(1):1-16. DOI: 10.1016/0304-4017(81)90002-9. View

Fitzgibbons P, Bradley L, Fatheree L, Alsabeh R, Fulton R, Goldsmith J . Principles of analytic validation of immunohistochemical assays: Guideline from the College of American Pathologists Pathology and Laboratory Quality Center. Arch Pathol Lab Med. 2014; 138(11):1432-43. DOI: 10.5858/arpa.2013-0610-CP. View

10.

Thomson S, Hamilton C, Hope J, Katzer F, Mabbott N, Morrison L . Bovine cryptosporidiosis: impact, host-parasite interaction and control strategies. Vet Res. 2017; 48(1):42. PMC: 5553596. DOI: 10.1186/s13567-017-0447-0. View

11.

Honda Y, Matsubara Y, Morzaria S, McKeever D . Immunohistochemical detection of parasite antigens in Theileria parva-infected bovine lymphocytes. Vet Parasitol. 1998; 80(2):137-47. DOI: 10.1016/s0304-4017(98)00184-8. View

12.

Pienaar R, Josemans A, Latif A, Mans B . The host-specificity of sp. (sable) and sp. (sable-like) in African Bovidae and detection of novel in antelope and giraffe. Parasitology. 2019; 147(2):213-224. PMC: 10317675. DOI: 10.1017/S003118201900132X. View

13.

Zweygarth E, Benade J, Steyl J, Prozesky L, Koekemoer O, Josemans A . In vitro cultivation of a Theileria species from a roan antelope (Hippotragus equinus). Parasitol Res. 2009; 105(6):1755-7. DOI: 10.1007/s00436-009-1589-4. View

14.

Hewitt S, Baskin D, Frevert C, Stahl W, Rosa-Molinar E . Controls for immunohistochemistry: the Histochemical Society's standards of practice for validation of immunohistochemical assays. J Histochem Cytochem. 2014; 62(10):693-7. PMC: 4212362. DOI: 10.1369/0022155414545224. View

15.

Ramos-Vara J, Miller M . When tissue antigens and antibodies get along: revisiting the technical aspects of immunohistochemistry--the red, brown, and blue technique. Vet Pathol. 2013; 51(1):42-87. DOI: 10.1177/0300985813505879. View

16.

Clift S, Steyl J, Mitchell E, Lawrence J, Hooijberg E . Clinical and hematologic features of experimental theileriosis in roan calves (Hippotragus equinus). Ticks Tick Borne Dis. 2021; 12(2):101644. DOI: 10.1016/j.ttbdis.2020.101644. View

17.

Schoch C, Ciufo S, Domrachev M, Hotton C, Kannan S, Khovanskaya R . NCBI Taxonomy: a comprehensive update on curation, resources and tools. Database (Oxford). 2020; 2020. PMC: 7408187. DOI: 10.1093/database/baaa062. View

18.

Nijhof A, Pillay V, Steyl J, Prozesky L, Stoltsz W, Lawrence J . Molecular characterization of Theileria species associated with mortality in four species of African antelopes. J Clin Microbiol. 2005; 43(12):5907-11. PMC: 1317158. DOI: 10.1128/JCM.43.12.5907-5911.2005. View

19.

Nene V, Kiara H, Lacasta A, Pelle R, Svitek N, Steinaa L . The biology of Theileria parva and control of East Coast fever - Current status and future trends. Ticks Tick Borne Dis. 2016; 7(4):549-64. DOI: 10.1016/j.ttbdis.2016.02.001. View

20.

Baker M . When antibodies mislead: the quest for validation. Nature. 2020; 585(7824):313-314. DOI: 10.1038/d41586-020-02549-1. View