Unit: Center for the Production and Infection of Anopheles

Director: Bourgouin Catherine

The platform CEPIA is mass-producing Anopheles gambiae and Anopheles stephensi, is providing lab facilities for infecting Anopheles mosquitoes with Plasmodium and for functional analysis in Anopheles (RNAI). The CEPIA has set up the in vitro production of P. falciparum gametocytes and mosquito infection with this parasite species. In 2005, 350 000 mosquito females were provided to research groups involved in mosquito-Plasmodium-vertebrate hosts interactions, mostly within the GPH Anopheles program.

The CEPIA, CEnter for the Production and Infection of Anopheles, is a platform the objectives and missions of which are organised around four main activities:

the production to the benefit of diverse research programs and teams, of Anopheles mosquitoes that are natural hosts and vectors or laboratory hosts for diverse Plasmodium species,

providing lab facilities, equipment and expertise for the research teams in order to:

a. perform mosquito infection by rodent malaria parasites,

b. perform mosquito dissections or manipulations

c. set up functional genomic studies in Anopheles, using tools such as transgenesis and RNAi

d. producing Plasmodium falciparum gametocytes and performing Anopheles infection with this parasite species

e. the development of collaborative projects that require Anopheles mosquitoes

Activity in 2005:

The moving of the Platform in its new setup since December 2004 allowed to :

   increase the mosquito production and strengthen its quality

   produce on a regular basis P. falciparum gametocytes, and initiate the production of P.falciparum-infected mosquitoes,

   to participate to the establishement of the RNAi tool in Anopheles sp. (Collaboration with the Unit "Biology and Genetics of Paludism")

1) Mass production of Anopheles mosquitoes and quality control

An. gambiae and An. stephensi have been mass-produced and distributed to 6 research Units within the Pasteur Institute (Biochimie et Biologie Moléculaire des Insectes, Biologie et Génétique du Paludisme, Immunophysiopathologie infectieuse, Réponses précoces aux Parasites et Immunopathologie, Génétique de la Différenciation, Parasitologie Biomédicale) and to a limited extent to teams outside the Institute. More than 350000 mosquito females were distributed, which corresponds to a weekly production of 7000 females and roughly 20.000 mosquitoes (Figure 1)

To verify that the produced mosquitoes have retain their full receptivity towards Plasmodium sp., a quality control approach has been set up and weekly infection are run with P. berghei on subsets of our mosquitoes

2) Production of Plasmodium falciparum gametocytes. Experimental infection of Anopheles with P. falciparum/

The ability to infect Anopheles mosquitoes with the human malaria parasite P.falciparum has two main goals :

first to facilitate molecular and cellular analyses of the interaction between the human parasite and a natural host and vector (An. gambiae), secondly to produce sporozoites, the parasite stage present in the mosquito salivary glands that initiates the early phase of the infection in the vertebrate host. Only the erythocytic development of the human malaria parasite is mastered in vitro. Therefore, the production of sporozoites through its mosquito host is required for any project aimed at understanding the biological processes (cellular, immunological) leading to the development of the sporozoites in humans.

Using an automated system for culturing, gametocytes from strain NF54 is produced on a regular basis. Those gametocytes are infectious to the mosquitoes and developed to the sporozoite stage (Figure 2). Experimental conditions for infecting Anopheles are under improvement for reaching a weekly production of P. falciparum infected mosquitoes.

3) Research programs

3-1 GPH "Anopheles"

The CEPIA is an essential partner for the GPH Anophèles (Grand Programme Horizontal) started in 2002. Beside the production of mosquitoes, the CEPIA participates in the development of tools for functional genomics (transgenesis and RNAi)

3-2 "Fonds dédié Special program" : Assessment of the contribution of An. gambiae salivary gland components to the infectivity of Plasmodium falciparum sporozoites ( C/ Mitri, I. Thiery, J.-C. Jacques, C. Bourgouin). The first part of this program initiated in June 2005 was aimed at optimizing conditions for reproducibly infecting An. gambiae with P. falciparum

3-3 Collaborations

The CEPIA is also participating in two other research programs developped at the IMTSSA (Dr C. Rogier) and the Paris University Pierre & Marie Curie (L. Lambrechts-J. Koella, E. Pondeville and C. Dauphin-Villemant) using Anopheles mosquitoes. Two other projects involved P. falciparum gametocytes (S. Briquet and C. Vaquero (Salpétrière) ; Linda Kohl (Museum National d'Histoire Naturelle)

4) Publications involving the CEPIA :

Demeure CE, et al, " Anopheles mosquito bites activate cutaneous mast cells leading to a local inflammatory response and lymph node hyperplasia." J Immunol. 2005 Apr 1;174(7):3932-40.

Lavazec, C., al. (2005). "cpbAg1 encodes an active carboxypeptidase B expressed in the midgut of Anopheles gambiae." Insect Molecular Biology 14(2): 163-174.

Figure legend : P. falciparum sporozoite labelled with fluorscent ant-CSP antibody /(Cliché Hannah Polson, on sporozoite produced by the CEPIA)

Keywords: Anopheles, production, Plasmodium, gametocyte, functional genomics

Activity Reports 2005 - Institut Pasteur

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