Comparison of Recombinant Proteins of Kinesin 39, Heat Shock Protein 70, Heat Shock Protein 83, and Glycoprotein 63 for Antibody Detection of Leishmania martiniquensis Infection

Abstract

Leishmania martiniquensis, a zoonotic hemoflagellate, is a causative agent of cutaneous (CL) and visceral leishmaniasis (VL) among humans and animals. This organism, first reported in Martinique Island, now has become an emerging infectious agent in Thailand. Symptomatic cases of L. martiniquensis infection among humans have continuously increased. In the meantime, asymptomatic infection of this novel species has seriously created national public health awareness and concern to prevent and control disease transmission. The unsuccessful serological test using the commercial rK39 dipstick based on antigen from Leishmania donovani to detect the antibodies against VL among infected Thai patients has encouraged us to further explore a new sensitive and specific antigenic epitope. In this study, we determined the sequences and expressed recombinant proteins of kinesin 39 (k39), heat shock protein 70 (hsp70), heat shock protein 83 (hsp83), and glycoprotein 63 (gp63) of L. martiniquensis to evaluate the diagnostic efficiency to detect antibodies against L. martiniquensis in patient sera. The preliminary results from western blot analysis have suggested that K39 is the most sensitive recombinant protein to detect L. martiniquensis. Moreover, this recombinant protein reacts with antibodies against L. donovani and Leishmania infantum, making it a promising antigen for further development of a universal rapid diagnostic tool for VL.

The rK39 Antigen from an Iranian Strain of Leishmania infantum: Detection of Anti-Leishmania Antibodies in Humans and Dogs

“…The k39 gene contains antigenic epitopes with amino acid variation affecting immunogenicity across Leishmania strains (20). The rk39 antigen that we used in the present study was prepared from an Iranian strain of L. infantum in the attempt to achieve the best diagnostic efficacy in Iran.…”

The increased presence of repetitive motifs in the KDDR-plus recombinant protein, a kinesin-derived antigen from Leishmania infantum, improves the diagnostic performance of serological tests for human and canine visceral leishmaniasis

“…One of the current challenges for the VL control revolves around improving the performance of diagnostic tests for human and veterinary use. Several studies have focused on the development of more accurate serodiagnosis, through replacing CSA by recombinant proteins [16,[37][38][39][40][41]. Until the early 1990s, the targets used in the serological tests were the crude or soluble antigens of promastigotes or amastigotes forms of Leishmania ssp., which has a high…”

Analysis of the Antigenic and Prophylactic Properties of the Leishmania Translation Initiation Factors eIF2 and eIF2B in Natural and Experimental Leishmaniasis

“…The intracellular location of the translation factors does not seem to be an impediment for their antigenicity. Similarly, many proteins with nuclear and cytosolic location, such as protein related to the translational machinery, enzymes implicated in parasite metabolism, heat shock proteins or histones have been described as immunodominant antigens not only in human and canine VL (Requena et al, 2000a ; Coelho et al, 2009 ; Soto et al, 2009 , 2015 ; Ramírez et al, 2013 ; Baharia et al, 2014 ; Sundar and Singh, 2014 ; Siripattanapipong et al, 2017 ), but also in human CL patients having moderate anti- Leishmania humoral responses (Ramírez et al, 2013 ; Souza et al, 2013 ; Duarte et al, 2015 ). Interaction of these proteins with B lymphocytes for antibody secretion may occur through complement mediated lysis of the non-metacyclic parasites causing the release of the whole internal cellular compounds (Mosser and Edelson, 1984 ; Ambrosio and De Messias-Reason, 2005 ; Moreno et al, 2010 ).…”