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*Leishmaniasis

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Clinical and Vaccine Immunology, February 2006, p. 271-276, Vol. 13, No. 2
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.2.271-276.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Kinetics and Diagnostic and Prognostic Potential of Quantitative Western Blot Analysis and Antigen-Specific Enzyme-Linked Immunosorbent Assay in Experimental Canine Leishmaniasis

D. Talmi-Frank,1 D. Strauss-Ayali,1,2 C. L. Jaffe,2 and G. Baneth1*

School of Veterinary Medicine, The Hebrew University, P.O. Box 12, Rehovot 76100, Israel,1 Kuvin Centre for the Study of Tropical and Infectious Diseases, Hebrew University-Hadassah Medical School, Jerusalem, Israel2

Received 28 August 2005/ Returned for modification 18 October 2005/ Accepted 1 December 2005

Quantitative computerized Western blot analysis of antibody responses during experimental canine Leishmania infantum infection distinguished between immunodominant and nonimmunodominant protein bands. Six infected beagles, positive by both PCR and parasite culture, were monitored over 75 weeks postinfection and during a 12-week allopurinol treatment course. All dogs were symptomatic at the time of treatment. Of 12 antigenic bands examined, the immunodominant bands (12, 14, 24, 29, 48, and 68 kDa) showed significantly increased intensities (P < 0.01) and higher frequencies of recognition than the nonimmunodominant bands at all time points. Detection of the former bands at 6 weeks postinfection preceded seroconversion by enzyme-linked immunosorbent assay (ELISA) both on crude Leishmania antigen or the recombinant proteins rK39 and HSP70. Reactivity with the 14-, 48-, and 68-kDa bands signified early infection, whereas increased reactivity with the 14-, 24-, and 29-kDa bands was associated with posttreatment parasite persistence and potential unfavorable prognosis. Total lane intensity (TLI) emerged as a sensitive marker for early infection and increased as early as 4 weeks postinfection. TLI had a significantly higher (P < 0.01) relative increase rate than crude Leishmania antigen or HSP70 or rK39 ELISA at all time points. These immunodominant antigens and TLI, as determined by quantitative Western blotting, will be valuable for early detection and treatment evaluation of canine leishmaniasis.

* Corresponding author. Mailing address: School of Veterinary Medicine, Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel. Phone: 972 39688557. Fax: 972 89489956. E-mail: baneth{at}agri.huji.ac.il.

Clinical and Vaccine Immunology, February 2006, p. 271-276, Vol. 13, No. 2
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.2.271-276.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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