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EFFECT OF MOSQUITO MIDGUT TRYPSIN ACTIVITY ON DENGUE-2 VIRUS INFECTION AND DISSEMINATION IN AEDES AEGYPTI

The effect of mosquito midgut trypsins in dengue serotype 2 flavivirus (DENV-2) infectivity to Aedes aegypti was studied. Addition of soybean trypsin inhibitor (STI) in a DENV-2 infectious blood meal resulted in a 91–97% decrease in midgut DENV-2 RNA copies (qRT-PCR analysis). STI treatment also resulted in slower DENV-2 replication in the midgut, less DENV-2 E protein expression, and decreased dissemination to the thorax and the head. A second uninfected blood meal, 7 days after the STI-treated infectious meal, significantly increased DENV-2 replication in the midgut and recovered oogenesis, suggesting that the lower viral infection caused by STI was in part due to a nutritional effect. Mosquitoes fed DENV-2 digested in vitro with bovine trypsin (before STI addition) exhibited a transient increase in midgut DENV-2 4 days postinfection. Blood digestion and possibly DENV-2 proteolytic processing, mediated by midgut trypsins, influence the rate of DENV-2 infection, replication, and dissemination in Ae. aegypti.

Received August 12, 2004. Accepted for publication October 20, 2004.

Acknowledgments: Funding for this research was provided from the National Institutes of Health, grant nos. R01 AI49256, R01AI45573, and U01AI45430. The authors thank the staff and students at AIDL for their assistance and helpful suggestions, especially Dr. Francisco Díaz and Dr. Irma Sánchez-Vargas.

Authors’ addresses: Alvaro Molina-Cruz (E-mail: amolina-cruz{at}niaid.nih.gov), Lalita Gupta (E-mail: lgupta{at}niaid.nih.gov), and Carolina Barillas-Mury (E-mail: cbarillas{at}niaid.nih.gov), Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Twinbrook III, MSC 8130, 12735 Twinbrook Parkway, Bethesda, MD 20892-8130, Telephone: (301) 435-2706, Fax: (301) 480-1337. Jason Richardson, Kristine Bennett, and William Black IV (E-mail: wcb4{at}colostate.edu), Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, Telephone: (970) 491-6136, Fax: 970-4911815.

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