Groupings of highly similar major surface protein (p44)-encoding paralogues: a potential index of genetic diversity amongst isolates of Anaplasma phagocytophilum

ABSTRACT Major surface protein 5 (Msp5) of Anaplasma marginale is highly conserved in the genus Anaplasma and the antigen used in a commercially available competitive enzyme-linked immunosorbent assay (cELISA) for serologic identification of cattle with anaplasmosis. This study analyzes the degrees of conservation of Msp5 among various isolates of Anaplasma phagocytophilum and the extent of serologic cross-reactivity between recombinant Msp5 (rMsp5) of Anaplasma marginale and A. phagocytophilum. The msp5 genes from various isolates of A. phagocytophilum were sequenced and compared. rMsp5 proteins of A. phagocytophilum and A. marginale were used separately in an indirect ELISA to detect cross-reactivity in serum samples from humans and dogs infected with A. phagocytophilum and cattle infected with A. marginale. Serum samples were also tested with a commercially available competitive ELISA that uses monoclonal antibody ANAF16C1. There were 100% sequence identities in the msp5 genes among all of the A. phagocytophilum isolates from the United States and a horse isolate from Sweden. Sheep isolates from Norway and dog isolates from Sweden were 99% identical to one another but differed in 17 base pairs from the United States isolates and the horse isolate. Serologic cross-reactivity was identified when serum samples from cattle infected with A. marginale were reacted with rMsp5 of A. phagocytophilum and when serum samples from humans and dogs infected with A. phagocytophilum were reacted with rMsp5 of A. marginale in an indirect-ELISA format. Serum samples from dogs or humans infected with A. phagocytophilum did not cross-react with rMsp5 of A. marginale when tested with the commercially available cELISA. These results suggest that rMsp5 of A. phagocytophilum is highly conserved among United States and European isolates and that serologic distinction between A. phagocytophilum and A. marginale infections cannot be accomplished if rMsp5 from either organism is used in an indirect ELISA.

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Cloning and expression of the gene encoding the major surface protein 5 (MSP5) of Anaplasma phagocytophilum and potential application for serodiagnosis

Veterinary Clinical Pathology ◽

10.1111/j.1939-165x.2006.tb00158.x ◽

2006 ◽

Vol 35 (4) ◽

pp. 418-425 ◽

Cited By ~ 5

Author(s):

A. Rick Alleman ◽

Anthony F. Barbet ◽

Heather L. Sorenson ◽

Nicole I. Strik ◽

Heather L. Wamsley ◽

...

Keyword(s):

Anaplasma Phagocytophilum ◽

Potential Application ◽

Surface Protein ◽

Cloning And Expression ◽

Gene Encoding ◽

Major Surface Protein ◽

Major Surface

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Molecular detection and genetic diversity of Anaplasma marginale based on the major surface protein genes in Thailand

Acta Tropica ◽

10.1016/j.actatropica.2020.105338 ◽

2020 ◽

Vol 205 ◽

pp. 105338 ◽

Cited By ~ 1

Author(s):

Witchuta Junsiri ◽

Amaya Watthanadirek ◽

Napassorn Poolsawat ◽

Sarawan Kaewmongkol ◽

Sathaporn Jittapalapong ◽

...

Keyword(s):

Genetic Diversity ◽

Molecular Detection ◽

Surface Protein ◽

Anaplasma Marginale ◽

Major Surface Protein ◽

Major Surface

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Differential Expression and Glycosylation of Anaplasma phagocytophilum Major Surface Protein 2 Paralogs during Cultivation in Sialyl Lewis x-Deficient Host Cells

Infection and Immunity ◽

10.1128/iai.01530-08 ◽

2009 ◽

Vol 77 (5) ◽

pp. 1746-1756 ◽

Cited By ~ 10

Author(s):

Matthew J. Troese ◽

Madhubanti Sarkar ◽

Nathan L. Galloway ◽

Rachael J. Thomas ◽

Sarah A. Kearns ◽

...

Keyword(s):

Differential Expression ◽

Cell Lines ◽

Anaplasma Phagocytophilum ◽

Surface Protein ◽

Host Cells ◽

Sialyl Lewis X ◽

Bacterial Populations ◽

Major Surface Protein ◽

Sialyl Lewis ◽

Major Surface

ABSTRACT Many microbial pathogens alter expression and/or posttranslational modifications of their surface proteins in response to dynamics within their host microenvironments to retain optimal interactions with their host cells and/or to evade the humoral immune response. Anaplasma phagocytophilum is an intragranulocytic bacterium that utilizes sialyl Lewis x (sLex)-modified P-selectin glycoprotein ligand 1 as a receptor for infecting myeloid cells. Bacterial populations that do not rely on this receptor can be obtained through cultivation in sLex-defective cell lines. A. phagocytophilum major surface protein 2 [Msp2(P44)] is encoded by members of a paralogous gene family and is speculated to play roles in host adaptation. We assessed the complement of Msp2(P44) paralogs expressed by A. phagocytophilum during infection of sLex-competent HL-60 cells and two HL-60 cell lines defective for sLex expression. Multiple Msp2(P44) and N-terminally truncated 25- to 27-kDa isoforms having various isoelectric points and electrophoretic mobilities were expressed in each cell line. The complement of expressed msp2(p44) paralogs and the glycosyl residues modifying Msp2(P44) varied considerably among bacterial populations recovered from sLex-competent and -deficient host cells. Thus, loss of host cell sLex expression coincided with both differential expression and glycosylation of A. phagocytophilum Msp2(P44). This reinforces the hypothesis that this bacterium is able to generate a large variety of surface-exposed molecules that could provide great antigenic diversity and result in multiple binding properties.

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Expression and Immunogenicity of Recombinant Immunoreactive Surface Protein 2 of Anaplasma phagocytophilum

Clinical and Vaccine Immunology ◽

10.1128/cvi.05709-11 ◽

2012 ◽

Vol 19 (6) ◽

pp. 919-923 ◽

Cited By ~ 2

Author(s):

Qiang Yu ◽

Chuang-fu Chen ◽

Qiang Chen ◽

Li-juan Zhang

Keyword(s):

Recombinant Protein ◽

Anaplasma Phagocytophilum ◽

Surface Protein ◽

Immunofluorescence Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Content Type ◽

Major Surface Protein ◽

Granulocytic Anaplasmosis ◽

Major Surface ◽

Diagnostic Reagent

ABSTRACTHuman granulocytic anaplasmosis (HGA), caused byAnaplasma phagocytophilum, is an emerging tick-borne zoonotic disease throughout the world. The first HGA cases in China were documented in 2008, and the greatest challenge posed by the disease is rapid and accurate diagnosis during the acute phage of illness. In this study, we successfully cloned and expressed anA. phagocytophilumimmunoreactive surface protein (major surface protein 2 [MSP2]) and demonstrated that this recombinant protein has natural immunogenicity by Western blotting and enzyme-linked immunosorbent assay (ELISA) using human HGA-positive sera and reference rabbit HGA-positive sera. The rabbit antisera against the recombinant protein also reacted actively with the natural antigen ofA. phagocytophilumby immunofluorescence assay (IFA). No cross-reaction was observed between the recombinant protein and rabbit antisera against 10 common members of the orderRickettsialesby ELISA when the sera were diluted more than 1:100. We concluded that the recombinant MSP2 protein exhibited excellent antigenicity and specificity, results that should lay the foundation for the development of a simple and rapid diagnostic reagent and a vaccination for anaplasmosis.

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CHARACTERIZATION OF HUMAN GRANULOCYTIC EHRLICHIOSIS IN EXPERIMENTAL INFECTED HL-60 CELLS WITH ANAPLASMA PHAGOCYTOPHILUM USING NESTED PCR AND A. PHAGOCYTOPHILUM MAJOR SURFACE PROTEIN-2 MONOCLONAL ANTIBODY

Bangladesh Journal of Veterinary Medicine ◽

10.3329/bjvm.v3i2.11400 ◽

2012 ◽

Vol 3 (2) ◽

pp. 160-165

Author(s):

MS Rahman ◽

JH Park ◽

JS Chae

Keyword(s):

Monoclonal Antibody ◽

Nested Pcr ◽

Anaplasma Phagocytophilum ◽

Surface Protein ◽

Intense Band ◽

Major Surface Protein ◽

Granulocytic Ehrlichiosis ◽

Human Granulocytic Ehrlichiosis ◽

Major Surface

The study was carried out to characterize the human granulocytic ehrlichiosis in experimental infected HL-60 cells with Anaplasma phagocytophilum using nested PCR and A. phagocytophilum major surface protein-2 monoclonal antibody. The nested PCR revealed only one band of 926 base pair DNA from A. phagocytophilum infected HL-60 cells. The western blot revealed several bands with a dominant 44-kDa. There were intense band of 100- and 160-kDa bands. The 44-kDa component was at least 10 times more abundant than the 100- and 160-kDa bands. In conclusion, the nested PCR would be a valuable tool for the characterization of human granulocytic ehrlichiosis.

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