Eappen G. Abraham

Johns Hopkins University

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Dana L. Vanlandingham

Kansas State University

Marcelo Jacobs‐Lorena

Johns Hopkins University

Sofia B. Pinto

Fundação Oswaldo Cruz

Kristin Michel

Kansas State University

Stephen Higgs

Kansas State University

Aidan Budd

European Molecular Biology Laboratory

Anil Kumar Ghosh

Oregon State University

Fotis C. Kafatos

Imperial College London

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Imperial College London

European Molecular Biology Laboratory

Johns Hopkins University

Kansas State University

Liverpool School of Tropical Medicine

University of California, Riverside

European Molecular Biology Laboratory

The University of Texas Medical Branch at Galveston

Centers for Disease Control and Prevention

Michigan State University

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An immune-responsive serpin, SRPN6, mediates mosquito defense against malaria parasites

Proceedings of the National Academy of Sciences (2005)

Eappen G. Abraham Sofia B. Pinto Anil Kumar Ghosh Dana L. Vanlandingham Aidan Budd

We have functionally analyzed the orthologous SRPN6 genes from Anopheles stephensi and Anopheles gambiae using phylogenetic, molecular, reverse genetic, and cell biological tools. The results strongly implicate SRPN6 in the innate immune response against Plasmodium . This gene belongs to a mosquito-specific gene cluster including three additional Anopheles serpins. SRPN6 expression is induced by Escherichia coli and both rodent and human malaria parasites. The gene is specifically expressed in midgut cells invaded by Plasmodium ookinetes and in circulating and attached hemocytes. Knockdown of SRPN6 expression by RNA interference in susceptible An. stephensi leads to substantially increased parasite numbers, whereas depletion in susceptible An. gambiae delays progression of parasite lysis without affecting the number of developing parasites. However, the An. gambiae SRPN6 knockdown increases the number of melanized parasites in the L3-5 refractory strain and in susceptible G3 mosquitoes depleted of CTL4 . These results indicate that AsSRPN6 is involved in the parasite-killing process, whereas AgSRPN6 acts on parasite clearance by inhibiting melanization and/or promoting parasite lysis. We propose that these observed phenotypic differences are due to changed roles of the respective target serine proteases in the two mosquito species.

Anopheles gambiae

Anopheles stephensi

Plasmodium (life cycle)

Prophenoloxidase

Serpin

Plasmodium berghei

10.1073/pnas.0508335102

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