FR2955393A1 - METHOD FOR ASSESSING THE EFFECTIVENESS OF VECTOR CONTROL STRATEGIES IN THE CONTROL OF MALARIA AND ARBOVIROSIS - Google Patents

Abstract

The subject of the invention is the use of immunogenic and specific salivary proteins of Anopheles and Aedes mosquitoes as biomarkers for evaluating the efficacy of anti-vector control strategies in the control of malaria and arboviroses, these proteins being chosen from the group comprising the proteins and peptides of sequences SEQ ID Nos. 1 to 30.

Description

Method for evaluating the effectiveness of vector control strategies in the control of malaria and arboviral diseases.

The subject of the invention is the use of immunological biomarkers to evaluate the efficacy of anti-vector control strategies (LAV abbreviated) in the control of malaria and arboviruses.

Parasitic and viral diseases are transmitted by arthropod vectors, such as mosquitoes in the case of malaria, dengue and chikungunya, ticks in Lyme disease or sandflies in leishmaniasis.

Among these diseases, malaria is the most serious parasitic infection in the world, the morbidity of which is directly related to human exposure to the infected Anopheles mosquito.

Many preventive methods have been proposed for the control of parasites (chemoprophylaxis) or their vectors (control of larvae or adults using larvicides and adulticides, insecticide impregnated supports, including mosquito nets, intradomiciliary sprays of insecticides and others).

The evaluation of the effectiveness of LAV strategies is based more generally on entomological methods for the vector, such as the evaluation of vector abundance, their aggressiveness, their infection rate or in humans on detection. pathogens. But these reference methods have many limitations in their ability to truly and individually evaluate the degree of pitting received by humans and thus the actual effectiveness of LAV measurements tested or used.

These limits are particularly encountered in a context of low exposure to vectors, which is becoming increasingly relevant in endemic and exposure areas.

Detection in humans, for example, of Plasmodium requires accurate and active monitoring of populations. The reference methods for evaluating LAV in the case of malaria most generally relate to the identification of Plasmodium in human blood (use of a thick drop). This method makes it possible to calculate the parasite prevalence and the intensity of infection which are tools used to evaluate the efficacy of LAV by comparing before / after introduction of LAV. Major limitations relate to the need for long-term population monitoring (over a year, for example) before and after LAV implementation. In fact, to say that LAV did reduce the prevalence or intensity of Plasmodium infection, it is necessary to make this evaluation taking into account the season of transmission. These limits are particularly important in areas where the transmission is seasonal or low (urban malaria, altitude) and where the distribution of antimalarial drugs is strong (failure to evaluate the effectiveness of a LAV against Anopheles by detection Plasmodium if antimalarial drugs are distributed to populations in the study area). In addition, the presence and intensity of infection of Plasmodium 30 can vary very regularly from one day to the next. As a result, it is very difficult to regularly monitor the populations of the area where LAV is assessed. This parasitological evaluation thus requires longitudinal follow-up studies in a large number of individuals to assert the efficacy of a LAV.

For arboviruses, the criteria for evaluating a LAV by detecting pathogens seem even more complex and not very usable. Indeed, the detection of an infection in humans is based on the production of IgM (infection in progress) or IgG (old infection) anti-pathogens. Aside from an epidemic situation, the prevalence of infection is often very low for arboviruses and the evaluation of an effective LAV therefore requires a very large study population before / after LAV to conclude that LAV is effective. Thus, for the evaluation of an anti-Aedes LAV, a very long-term follow-up is necessary and requires very heavy and expensive field logistical means.

In addition, entomological methods are mainly applicable at the level of a population, but do not give a measure of the heterogeneity of the individual exposure.

Thus, the interest for new indicators and methods will be measured in order to evaluate at the individual level, the real human-vector contact and thus the real effectiveness of a LAV strategy, in conformity with the recommendations of the WHO.

A coordinated research program in a laboratory of the Depositor focuses on the immunological study of the human-vector relationship during malaria and arboviroses.

Previous work has shown that antibody responses specific to saliva total or specific salivary protein extracts of Anopheles and Aedes represent relevant biomarkers for assessing the degree of exposure of human populations to mosquito bites. More specifically, the research group to which the inventors belong has demonstrated that the total anti-salivary IgG antibody response of An. gambiae (1) and (2) and anti-peptide of the gSG6 protein of this Anopheles vector (3) was a tool for assessing exposure to the malaria vector. In addition, the total anti-salivary Ig antibody response of Anopheles gambiae was also a tool for evaluating the efficacy of insecticide-treated bed nets (hereinafter abbreviated as ITN for "Insecticide"). Treated Nets ") (5). Immunogenic salivary proteins constituting potential candidates as biomarkers of exposure to stings have been identified.

The validity of this approach has also been demonstrated for the development of a tsetse fly biomarker, vector of THA (4) and (3).

As part of the continuing work in this area, the inventors have found that immunoepidemiological markers constituting diagnostic indicators of malaria risks and markers of exposure to the bite of this vector, were also specific biomarkers for assessing the risk of malaria. effectiveness of a LAV.

The invention relies on the use of such immunologic biomarkers to evaluate the efficacy of an LAV strategy.

It is also an object of the invention to provide a method for evaluating the efficacy of an LAV strategy based on measuring antibody levels against salivary proteins and peptides, specific biomarkers of exposure to Anopheles mosquito bites and Aedes.

It is particularly aimed at the application of this method for anti-vector control aimed at reducing the transmission of malaria and major arboviruses. According to a first aspect, the invention relates to the use of immunogenic and specific salivary proteins of Anopheles and Aedes mosquitoes as biomarkers for evaluating the efficacy of vector control strategies in the control of malaria and arboviroses, these proteins being selected from the group comprising the proteins and peptides of SEQ ID NO: 1 to 30 given below.

According to a second aspect, the invention relates to the use of the human antibody response to immunogenic and specific salivary proteins of the Anopheles and Aedes mosquitoes of said sequences SEQ ID Nos. 1 to 30 for evaluating the efficacy of vector control strategies. in the control of malaria and arboviruses. According to another aspect, the invention also provides a method for evaluating the efficacy of an LAV strategy in the control of malaria and arboviroses, characterized in that it comprises the evaluation of the specific antibody responses to immunogenic salivary proteins and / or peptides of Anopheles and Aedes mosquitoes as a criterion for efficacy of LAV, regardless of the larvicidal or adulticidal form of LAV, these proteins and peptides being selected from the group consisting of proteins or peptides corresponding to SEQ sequences ID Nos. 1 to 30.5 The sequences SEQ ID Nos. 1 to 30 mentioned above are as follows:

SEQ ID NO: 1: gSG6 protein with the signal peptide underlined.

MAIRVELLLAMVLLPLLLLESVVPHAAAEKVWVDRDNVYCGHLDCTRVATFKGERFCTLCDT RHFCECKETREPLPYMYACPGTEPCQSSDRLGSCSKSMHDVLCDRIDQAFLEQ

SEQ ID NO: 2 p1 peptide of the gSG6 protein, hereinafter designated gSG6-P1 EKVWVDRDNVYCGHLDCTRVATF

SEQ ID No. 3: peptide P2 of the gSG6 protein, hereinafter designated by gSG6-P2 ATFKGERFCTLCDTRHFCECKETREPL SEQ ID No. 4: peptide P3 of the gSG6 protein, hereinafter designated gSG6-P3 CECKETREPLPYMYACPGTEP

SEQ ID NO: 5: P4 peptide of the gSG6 protein, hereinafter designated gSG6-P4 ACPGTEPCQSSDRLGSCSKS

SEQ ID No. 6: P5 peptide of gSG6 protein, hereinafter referred to as gSG6-P5 DRLGSCSKSMHDVLCDRIDQAFLEQ

SEQ ID NO: 7: protein gVAG MAIWIACATLLLVVLSGVSAGGKYCSSDLCPRGGPHVGCNPPSSSGGPTCQGKQKARKVLLT 30 PALQAYIMDEHNLNRSNIALGRIRPYPSAVKMPTLTWDPELASLADANARSCNYGHDRCRAT KKFPYAGQNIAITQFFGYRFTEKDLIHKFVSSWWSEYLDARPEHVRKYPSSYSGKPIGHFTQ IASDRSTKVGCSMWYWKDGQMDVYYFVCNYSFTNIMDRSVYQSGPTASQCKTGRNSKFPGLC NASEEPRSIMDP15 SEQ ID NO: 8 Protein 5 'nucleotidase MALVRFATIITVLCHLAIQDGAAKSFPHGEKAPFPLTLIHINDLHARFDETNQKSSTCTNSK ECIAGIARVYHTIKQLKSEYKTKNPLYLNAGDNFQGTLWYNLLRWNVTAYFIKELPPDAMTL GNHEFDHSPKGLAPYLAELEKMKIPTVVANLEKNGEPALKDSKIAPQIVLKVGNRKVGVIGA LYDKTHLVAQTGMVTLTNSIEAVRKEAQELKKKNVNIIVVLSHCGLDGDKQLAEEAGDLIDV IVGAHSHSLLLNKDAKVPYDTKYDTIEGDYPLVVKKSNNHTVLITQARSFGKYVGRLTVNFD CEGEVQSWEGYPIYMNNSVKQDEEVLRELEPWRAEVKRLGTQVIGTTEVFLDRESCRWCECT LGDLIADAYADQYTNSTVQPVAFVQAGNFRNPIEKGDITNGLAIEAAPYGSSVDMIKLSGAD LWSAIDHSFTLDDEFRYNTAQVSGMTVVADLSKKPYERVQSIDIVGANGAKTALKKDQIYYV AVPSYLADGKDGFAMLKKGTNRVTGPLDSDVLIEYVRKRQTIAKTMFQEKRVVIENHTNGTC SWDLDSQRYKPK

SEQ ID NO: 9 SG1 protein-like 3 MAGQRHLIEQAWQYGAQLQHELMLTSMESDRVQRALVLHSMLVNASLAEMVKESYQTHGADG RMVVRMLKFVRLLPGADERVAVYKQLAELLKSNGQDGRFPAVIFSTDVRQLEDRYKPDHAQY EGKVVERWLAELQAGTFHEVVEFARDYPEYFARVEEPLYETLKQQWSAEGLDRMVSFPNALP VGVQRVRALRALLETLLQHQGEQNNDVYLIRLAHETGRVEATVGQADAAVRQALDDVKKLFE QFKYQRGFPDYEALYKLFKGL SEQ ID NO: 10: A protein 34 kDa 1, with signal peptide (underlined) MSPSNKILVLLLFPILLVSSHPIPAEDPAKQCNLSEDDLTKLKAAISGASSAKAANEDILPN TTLAACPMLKNFTEMLKTVATDMEVLKTQGVSNMEVQLLRESFEEKLNDLAKNKDIFERQAN QDTSKAEGEMVEKINKLQLEMAKLQEEIEEQTKQMYVDMIEYIFERLKMNDTEAIDSYAQIV MKTKMHELIMKLKTDRLVLWEMVKYVEGKKNKWVGRKVLNTILDQVNKLKLYKPEEVEIGKN SLVVVWCWKFNSETVYGTTDEDQKSFHLAKLFFPKEKGCKECANVKSRTMCNNDYPKVMVKA FG

SEQ ID NO: 11: A protein 34 kDa # 2, with the signal peptide (underlined) METSLPITVVFPIVLITGAQTKPTQGSCTLTDEDISDIKSAVQKASKAAVNDIVLDPTLIDK CPMLEKITASLKSVATEIVQMRDSAISTDQVDQLKQNFEDQVNQIVKSRDIFEKQSGTQATK EHGEMLERMTAQVKVTELEQQIAKQTASMYEDMAELIFQRLQMNSTESVRSYTKHMMEEKLE ELMNKLETNYRIYLGALRFLNHMNDQELIGKVFDGILKRLGDMKADSDDVKENGRNLLVNLL CWTVNNDFLGKKYKERQVDLYRMALKFYPKTYEKAANEADVRSRQFCEENFPANLITWFAVS WNDRG

SEQ ID No. 12: 34 kDa Protein No. 1, with Signal Peptide (underlined) The biomarker peptides are indicated in italics. MKTSLPIVVLLTAVISGVHPNPTPKSCTVSEEDLTTIRNAIQKASRASLDDVNLDEDLIAKC PLLKTITASLKSVASEIATLKDTGISEEQVDELKQSYEQQVNEIVKSRDIFEKQSGGDVMKE QGAMINRMTELQVQVAQLQQQIGEQTSRMYDDMAELIFQRLAMNSTDSIRNYTAHMMEQKLH TLMTKLETNYRIFLGALRYLDHLGDQPLIDKVFDGILKRLDEMSLETNKERENGKYVLVNLL CWTVNNRFLTEKYRKKQLELFRIALKFYPKTGNKEANEADIRGRQFCDANFPVNVITWFAVS RAAEGWGLRGTL

SEQ ID NO: 13: A protein 34 kDa # 2, with the signal peptide (underlined) Biomarkers peptides are printed in italics MPVTYEFFILLAMSLLLVRSHPLPEEATSDAAIKCTLSEEDLDSLKRAISSAASAKSSDGII LSNETLTACPILANFTEMLKTVATDMEVLKTQGVSNAEVELLRESFEERLNEITKNKDIFER QAGQETSKTEGQMVEKINRLQLEMASLKEEIEQETKKMYEDMAEYIFQRLKMNDTDAIDSYA QIMFKTKMHDLFMKLKTDRWVLWKMLNYVEQKKDKVIGKWVLNTVINQVTSLNRNKPDELEI GKDSLVNLLCWTSTAKTVYGAVQEDQKMFYLT

SEQ ID NO 14: Peptide N-Term of the protein 34 kDa HPIPAEDPAKQCNLS SEQ ID NO: 15: A protein 62 kDa # 1 MNPFTQVFCLIVGVLFFAFTATADEDCEIPLSELNKIEQTLTHIEKPIYTEEAESETSDSDE CAQMLRGIHLQLRRLTQKYKLMNKGYVKVEEFHKMAKEYEEQLSKLNGELEYFKVEARSGAD QKIKELKKDIKSLEQNVTKLHDRLKGITDELGKVSMELCSTYLESNQLDSAKEKVKDLPSKY VIELVDQFLSKNENNWLPVVDLSTAVPDLDDRGQVYKNIYEFLLAKSRLEGEESLLLEAEIL KLNATFHPGSKITDDRKKELNDLLEKLRQNSEKTFEQWSQELDQSNISAVVFKNAIDRMFLT IQ EKFGEYVTGRHDYYGVRNFLKLLGVSKNYHKIAAYNSLVEKKIGHALAVVMIDMLSIDIA EIKHDPHVPDRIERMYNSTINSLPDSLKGIIPCIKSVKIRNEGTNRCIYAINQNIQVQNPNF KNIVLGQYKRVTSTMSACTSFRLELSSNKPYIKIITSEGNSLTNINSAVPGETGFSRVGAPY SNKPNMKLDHTGDWVLDANFVNNSIKIQSDFNAYQTSKSIDHLVVRNDGDVAVVRYGLSGMR SGGIPDNHAEWKF

SEQ ID NO: 16: A protein 62 kDa # 2 MNPFTQVFRLTVGVFFFAFTATADEVCEIPLSELNKIEQTLTHIEKPIYTEEAESETSDSDE CAQMLRGIHLQLRRLTQKYKLMNKGYVKVEEFHKMAKEYEEQLSKLNGELEHFKVEARSGAD QKIKELKKDIQALEQNVTKLHDRLKGITDELGKVSMELCSTYLESNQLDSAKEKVKDLPSKY VIELVDQFLSKNENNWLPVVDLSTAVPDLDDRGQVYKNIYEFLLAKSRLEGEESLLLEAEIL KLNATFHPGSKITDDRKRELNVLLTKLSQFSEKTLEQWTQELNQSNISAVVFKNAIDRMFLT QIEKFGEYVTGRHDYYGVRNFLKLLGVSKNYHKIAAYNSLVEKKIGHALAVVMIDMLSIDIA EIKHDPHVPDRIERMYNSTINSLPDSLKGIIPCIKSVKIRNEGTNRCIYAINQNIQVQNPNF KNIVLGQYKRVTSTMSACTSFRLELSSNKPYIKIITSEGNSLTNINSAVPGETGFSRVGAPY SNKPNMKLDHTGDWVLDANFVNNSIKIQSDFNAYQTSKSIDHLVVRNDGDVAVVRYGLSGMR SGGIPDNHAEWKLNALK SEQ ID No. 17: 62-kDa Protein 1 MNSVNRYVLCIALIALFVASFTTAEENCEISANELGKIEQTLTHINQPIYTGDDESEVSDSD QCAQMLRGIHFQLRRLTQKYKLMNKGYVKAEEFAKMARDYEDQLSVLKNDLEQSKIGADSSA KQKMQELKKDIATLEQNVNTLHKDLEGITDELGKVRMDLCLTYMESNQLSNAQDKVKTLAPK YLMELVEQFLNKSEKNWLPVVDLSVAIPDLDDRGQVYKTVHEFLKTKNRDGGEDSILLEAEV LKMNATFHPGSKITEDRKKEIQDLLEKLSLTSTKIFDQWTQDLAKLENSAVYKNSIDRMFLT QMEKFGERVMAKDDYYSLRNFLKLLVVS TNYYKIAAYRKLIQEKIGHTLAVLMFDMMSMERT ELQYDPHVPDEVVRMYDESITALPDSLKNIRSCLKLVKIYNHVTNQCILATNEVEDVNNSNP KFKSNVLGRRKLVKTASNDCTPFRLEPSADKASIRIITPKGDALTNINSIQPGLSWFNRVGA PYTNNHNMKLDYSADWILDANYANDSIKIESEFNAYQTMKSVDHLMVTNVGKVPHVVVAQYG LKGMEYAGAGMKDAEWKFKCDN

SEQ ID NO: 18: A protein 62 kDa # 2 MKVYICQVIFSFLAVSVFCEENCNIPESELSKIDHVLRHMEKPIYSEEQFASDNEECTNLLN GIHAQLRRLTQRYKLMNKGYVKVEEYQRMADNYEKQLKTLNDELVELQQHTSEKASATIAKL KEDIKKLDEEVGTLHEKLKGIKQDFEKVKRDLCVTYLNSNQMSKAKAKLKEMASTYLIEIVQ QQLNKSNANIMPMLEFSAAIPDLDDMGEAYKEIYKFLEEQKRLEGEDSVLLEATVLKMNASL KEGSNITDERRTQIEGLLKDLATKSTIVFSTWTKELKKINDAVVIKNALDHMFVSQMKVFGA LVGDTSDFGSIRNFVKLTIVCNNYYKVAAYKELIDRKIGNALGTIMFDLLTLEVNEMKFDPH VPDEIPKLFEATLSSLPNSLTELRTCLGKVQIYNKKTNKCVVATGNDFDVHKDKLGDFYRVV VADYGCTSFRLEASGDKASVRIVTPSGNPMSNVNLHLEGNSLHNYVATPKSNKPDRTPSSSD EWILDANYNNDTIKIESQFSDYKTKKTEVDHLLVRDINHLPHVLVARYGFMGLKNSDAKDTI EWNLKCGS SEQ ID NO: 19: A protein 62 kDa # 3 MKWSVYIALLVFAFLTSPVFSEENCNIPESELSKIDDVLRHMEKPIYSEDHYTSNNEECTNL LNGIHAQLRRLTQRYKLMNKGYVKVEEYKRMAEDYENQLKTLNAELLELQEHTSDKANAAIA KLKEDIKKLDEDVDTLHNKLKGIKQDFEKVKRDLCLTYLNSNQMSNAKAKVKEMASTYLIEI IQQRLNTKYANIIPMLDFSTAIPDLDDRGEAYKEIYKFIETHERLDGEDAVLLEASLLKMNA TLKEGSNITDERRTEIEKMLKELAEKSAVVFKTWSTELKGIEDTIIKYALDHLFVNQMKVFG GIVGDTFEFAPIRHLLKLLVVCNNYYKVAAYKELIDR KIGNVLGTIMFDLTTLEANEMSFDL HVPDEIPKLFNATLGSLPNSLTQLLPCLNKVHVYNAKTNMCIVAPEDRFDVQQEKLTDFHRV VLAKYGCTAFRLESSPNKASVKFVKPSGNALSSINLQLENDQWHSHVGTPTANKPDRKPSSS DEWILDANYVNDTVKIQSEFNEYKASQAEVDHLLVMDVKYLPHVVVGRYGVRGLKRSSAKDT IEWYLKCAS

SEQ ID NO: 20: Long Protein D7 form # 1 MKLPLLLAIVTTFSVVASTGPFDPEEMLFTFTRCMEDNLEDGPNRLPMLAKWKEWINEPVDS PATQCFGKCVLVRTGLYDPVAQKFDASVIQEQFKAYPSLGEKSKVEAYANAVQQLPSTNNDC AAVFKAYDPVHKAHKDTSKNLFHGNKELTKGLYEKLGKDIRQKKQSYFEFCENKYYPAGSDK RQQLCKIRQYTVLDDALFKEHTDCVMKGIRYITKNNELDAEEVKRDFMQVNKDTKALEKVLN DCKSKEPSNAGEKSWHYYKCLVESSVKDDFKEAFDYREVRSQIYAFNLPKKQVYSKPAVQSQ VMEIDGKQCPQ SEQ ID NO: 21: Long Protein D7 form # 1 MNILLVLAVVTISSLALVTAKGPFDPEEMHFIFTRCMEDNLKDGPDRVKTLLKWKEWVTEPK DDPATHCFAKCVLEMSGLYDAASGKFDASVIEAQHKAYPNSEDKGKVDAFVKAVQALPPTKN DCTAVFRAFGPVHMAHKATSINLFHDNKALTKGIYEKLGKDIRQRKQSYFEFCENKHYPVGS PKRSDLCKIRQYVVLDDAQFKQHTDCIMKGLRYITKDNILNCDEIKRDFKQVNKDTGALEKV LNTCKAKEPRDVKEKSWHYYKCLVESSVANDFKEAFDYREVRSQNYGYHLMQKQPYNKPAVQ AQVSEVDGKQCPS SEQ ID NO: 22: A protein 30 kDa (allergen) # 1 MKPLVKLFLLFCLVGIVLSRPMPEDEEPVAEGGDDDASGESEGEEETTDDAGGDGGEEENEG EEHAGDKDAGGEDTGKEENTGHDDAGEEDAGEEDAGEEDAGEEDAGEEDAEKEEGEKEDAGD DAGSDDGEEDSTGGDEGEDNAEDSKGSEKNDPADTYRQVVALLDKDTKVDHIQSEYLRSALN NDLQSEVRVPVVEAI GRIGDYSKIQGCFKSMGKDVKKVISEEEKKFKSCMSKKKSEYQCSED SFAAAKSKLSPITSKIKSCVSSKR

SEQ ID NO: 23: A protein 30 kDa (allergen) 2 MKPLVKLFLLFCLVGIVLSRPMPEDEEPVAEGGDEETTDDAGGDGGEEENEGEEHAGDEDAG GEDTGKEENTGHEDAGEEDAGEEDAGEEDAGEEDAEKEEGEKEDAGDDAGSDDGEEDSTGGD EGEANAEDSKGSEKNDPADTYRQVVALLDKDTKVDHIQSEYLRSALNNDLQSEVRVPVVEAI GRIGDYSKIQGCFKSMGKDVKKVISEEEKKFKSCMSKKKSEYQCSEDSFAAAKSKLSPITSK IKSCVSSKGR SEQ ID NO: 24: A protein 30 kDa allergen # 3 MKYLLTFLMALSLVNLMLTRPTPEDDGGTSEEPQTQETTGSDEKNGASEEPNADDASKPDDV EEKGDDDTAKKEDDGESKDGEGSEKSDKEKGEPKNDPRETYNKVIEQLDQIKVDNVEDGHER SELAADIQRYLRNPIVDVIGSAGDFSKIAKCFKSMVGDAKKAIEEDVKGFKECTAKKDSNAY QCSQDRSTVQDKIAKMSSKIASCVASNRS SEQ ID NO: 25: A protein 30 kDa allergen n ° 1 MQLFPISLVVSCLVFSFVLSLPLPEEEASTSEETEATTEAESAAADETTATGADDAESGSSE KNNEGGETSAPEAKEEGSGKDDRLDTYNKVNELLDKGTGVNNVENGFLRAFLDNNLQSHLRV PVIDAIGLVGDFSKIAGCFTSMGADVKKIVDDKLKSFTTCKSGKEAGDAKCSEDGSSVGGQF DQITSKIKECVSSKGKGK

SEQ ID NO: 26 A protein 30 kDa, Allergen 2 MRSLLVLFAALCLVGFVLAKPIPEDDTEATTEEPKAEDAAEQTTAEDAGGSDGEKNDDEKPS DDAASNDQPKDVEGGEDGKESPKTEGEAKNDPQETYNKVIELLDQIKVDNVENGYERSELTS DLQTYLRNPIVDAITVGDFSKIADCFKSMGDEAKKAIEEELKAFKECTDKKESDAYQCSKNH STVQGKIAKISSTINSCVVSKRA SEQ ID NO: 27: A protein apyrase / 5'nucleotidase No. l MARLSFVPLVISFLSCTTALSEKSLFPLSIIHVNDIHARFEETNYVGTVCKPHEGQVCIGGY ARTVTVVKHLLRIRQNPLYLNAGDNFQGTLWYNMFRWNVTSTFLNMLPADAMTLGNHEFDNG VDGVIPFLDTASSPVLLCNVDASEVPEFVKYEKSIVIERAGRKIGIIGVILKNTSVISNPGK LRFLDEASSVKSEAQILKNQGIDIIVVLSHCGVDVDRRIATEGGEHVDIIVGGHSHTFLYSG NNTGIPGTVQGSYPTVVTHASGHKVLIVHAAAYTKLVGDIVLYFDEQGIIQRWEGNPHYLDP EVVPDPEVHKALLPWKLAVDELGNRIIGTTITDLPRLGCNYRECALGNLISDAFVDSSAYMA EPGHWSYASIGLTNTGGIRVGLSKGDISYKSLIEVIPFENTVDFIELRGDHLLRVLEHGTAK SNIQISGLRVVYNMTEPYGNRVKSVQARCHGCSEPRFDPLDPFKYYRVAVNSHMAGGGGGFT MFEEFGRNKVVGDVEINALVRFVQKRSPLRYEVEGRVTVLN

SEQ ID NO: 28 Protein apyrase / 5'nucleotidase # 2 MARLSFVLLVLLFLTYMTVIAEKPLFPLSIIHVNDIHARFEETNYVGTVCKPHEGQVCIGGY ARTVTVVKHLLRIRQNPLYLNAGDNFQGTLWYNMFRWNVTSTFLNMLPADAMTLGNHEFDNG VDGVIPFLDTVSSPVLLCNVDASEEPEFVKYEKSIVIERAGRKIGIIGVILKNTSVISNPGK LRFLDEASSVKSEAHILKDQGIDIIVVLSHCGVDVDRRIATEGGEHVDIIVGGHSHTFLYSG NNTGIPGTVQGSYPIVVTHASGHKVLIVQAAAYTKLVGDIVLYFDEQGIIQRWEGNPHYLDP EVVPDPEVHKALLPWKLAVDELGNRIIGTTMTGLPRLNCNYRECALGNLISDAFVDSSAYMA EPGHWSYASIGLTNTGGIRVGLSKGDISYKSLIEVIPFENTVDFIELRGDHLLRVLEHGTAK SNIQISGLRVVYNMTEPYGNRVKSVQARCHDCSEPRFDPLDPFKYYRVAVNSHMAGGGGGFT MFEEFGRNKVVGDVEINALVRFVQKRSPLRYEVEGRVTVLN

SEQ ID NO: 29: A protein apyrase / 5'nucleotidase # 3 MLRTSSIVFLTCCLTFLIEGSSFKLKIIHFNDIHARFDEVTNSSSPCSGNGETCVAGIARLV TTIEKLRKQNENHLVLNAGDVFQGTIWYTLLKWNVSQQFMNMVKADAMTLGNHEFDDSFPVL IPFLENTKNVTPVVVSNLVFPKQLSRDVTKFRSLIKEDPLVLTVGGQSIGIIGVIFDETDKI GNADPLKFKSSIETVRIAAKQLKSKGVNIIIVLSHCGVFDDKKIAEQAGEDIDIIVGGHTHT LLYNGDPPSKHAALDKYPIVVETGNNHKVLIVQAFCHGHYVGNIDLTFDDEGEITAFEGQPI YQENRIEKNALVEARVRELRKDVEVKSLVKVGESKLELSNDCRLKDCTFGSVLADAYVWHFR SRSNAPMIAMIHPGNFRISLAAGVITRGQILTALPFNSNANRVTVLGSTIKKAIEFGTSINP RRCSFNALQTAGIKIDVDYGKPVGNRTVILLKTGGKYKRLVESKKYDILVNSYVFKGGDGFD MFKHLAVKGRAPFDAELLEQYIVARKGIQKSGLLQSRMNVSHVEKALSEVKSCKQR SEQ ID NO: 30: A protein apyrase / 5'nucleotidase # 1 MAGKPGIQLFVIFILLSSFAAVVWTTDNMPADKDVSKLFPLTLIHINDLHARFDETNMKSNA CTAKDQCIAGIARVYQKIQDLLKEYKSKNAIYLNAGDNFQGTLWYNLLRWQVTADFITKLKP TAMTLGNHEFDHTPKGLAPYLAELDKAGIPTLVANLVMNDDPDLKSSKIQKSIKVTVGGKTI GIIGVLYDKTHEIAQTGKVTLSNAVETVKREAAALKKDKVDIIVVLSHCSYDEDKKIAKEAG QDIDVIVGAHSHSFLYSKESNKPYDQKDKIEGPYPTIVESNNKRKIPIVQAKSFGKYVGRLT LYFDNEGEVKHWEGYP EFIDNKVKQDPKILEALIPWRKKVQEIGSTKVGETTIELDRDSCRD KECTLGVLYADAFADHYTNSSFRPFAIIQAGNFRNPIKVGKITNGDIIEAAPFGSTADLIRL KGDSLWAVAEHSFALDDENRTNCLQVSGLRIVIDPSKKIGSRVVKIDVMDNRNPKSEDLKPL DKNAEYFIALPSYLADGKDGFSAMKKATARWTGPLDSDVFKSYVEKIKKVDKLKWGRVIVCK AGSPCT

These immunogenic salivary proteins and / or peptides derived from these immunogenic proteins have been identified as immunogenic and Anopheles or Aedes-specific proteins and peptides and as vector puncture exposure biomarkers.

In accordance with the invention, these proteins and peptides have been selected as biomarkers of efficacy of LAV.

Specific biomarkers more particularly correspond to the sequences SEQ ID Nos. 2, 7, 9, 10, 12 and 14.

The antibody responses specific to these proteins and peptides represent a criterion for evaluating the efficacy or non-efficacy of LAV strategies against Anopheles and Aedes mosquitoes as illustrated by the examples. It is more particularly the IgG antibody and isotype (IgG1, IgG2, IgG3, IgG4, IgM, IgE and IgA) responses.

The method according to the invention is characterized in that it comprises the evaluation of antibody responses specific to these salivary proteins as a criterion for efficacy of LAV, regardless of the larvicidal or adulticidal form of LAV.

Targeted Anopheles mosquitoes include Anopheles gambiae and Anopheles funestus complexes. Aedes mosquitoes include Aedes aegypti and Aedes albopictus.

The sequences SEQ ID Nos. 1 to 30 above correspond more particularly to proteins or peptides isolated from salivary extracts of Anopheles or Aedes, namely, for -SEQ ID No. 1-9, Anopheles gambiae and optionally Anopheles funestus, SEQ ID NO: 1, 11, 27-30, Ae.aegypti-SEQ ID NO: 12, 13, 15, 16, 25, 26 and 30, d Ae. Albopictus.

Surprisingly, the peptide P1 of the gSG6 protein of sequence SEQ ID No. 2 is a biomarker of low or very low exposure to the Anopheles vector (6). Under such conditions, entomological methods would have very high exposure assessment limits. However, according to the invention, this specific biomarker has proved particularly suitable for the evaluation of LAV which greatly reduces mosquito density if it is effective.

The Anopheles total anti-saliva IgG response thus represents a new immunoepidemiological and individual indicator for evaluating the efficacy of anti-malaria vector strategies, such as the use of insecticide-treated mosquito nets and the like. LAV forms by larvicides and adulticides.

For Aedes, Ae.aegypti's total IgE and IgG4 responses to total saliva increased sharply during the puncture exposure period of this vector (7), and IgM and IgG anti-saliva responses can detect Aedes exposure to Aedes. travelers in endemic areas for arboviruses (8).

The invention thus makes it possible to evaluate, in the short term, the effectiveness of new LAV strategies for Anopheles and Aedes, for example within about 6 weeks after their introduction. It also allows longer-term monitoring (for example over a year) of the LAV used, thus representing an alert to indicate that the LAV used is not effective over time.

The invention also makes it possible to evaluate the real impact of LAV on a decrease in human-vector contact, hence the real impact of LAV on the risk of transmission of pathogens.

Target populations include those living in endemic areas as well as travelers.

Advantageously, the evaluation of LAV according to the invention can be measured in a small number of individuals, more particularly in a sub-sample of the total population concerned by the introduction of LAV.

It is also interesting to compare the effectiveness of different LAV for example larvicides versus adulticides, 30 mosquito nets versus insecticide sprays.

Other characteristics and advantages of the invention are given in the following examples in which reference is made to FIGS. 1 to 17.

These figures represent, respectively: FIG. 1, the evolution of the IgG anti-gSG6-P1 response during the follow-up period (before and after introduction of 5 long-lasting insecticidal nets, hereinafter LLIN in abstract); - Figure 2, the impact of LLINs on the decline of anti-gSG6-Pl IgG responses; - Figure 3, the number of responders according to the 10 months before and after LLINs; FIG. 4, the profile of the anti-P1 IgG responses of the responders; - Figure 5, the comparative evolution of the average densities of An. gambiae and the median IgG anti-P1 response; Figure 6 shows the comparative evolution of mean parasite densities and the median anti-P1 IgG response; - Figures 7-9, the evolution of IgG anti-gSG6 P1 response in 0-6 years, in 7-14 years, and in +14 years; Figure 10 shows the comparative evolution of the IgG anti-gSG6-P1 response in different age groups; 11A-11C, the profiles of the individual anti-Pl IgG responses before and after LLINs in different age groups; - Figure 12, the average, median and extended distributions in different age groups for each passage; - Figures 13 and 14, the April responder profiles in July and August 2006 (Fig.13) and July in April and August (Fig 14); - Figures 14 and 16, the age distribution of the answering machines of June-July 2006 and the numbers according to the age of the answering machines of June-July; 17, the comparative evolution of the anti-gSG6 -P1 IgG response by sex before and after LLINs.

Example 1 Validation of the Pl peptide of the gSG6 protein of An. gambiae, of sequence SEQ ID No. 2, as a marker for low exposure to the bite and as a tool for evaluating the effectiveness of LLIN treated nets.

The results of a study conducted in the locality of Lobito (West Uganda) on 105 individuals from 19 families, before and after LLINs are used. The most important malaria vector in this region is An. gambiae sl

1- Evolution of the anti-gSG6-P1 IgG response during the study.

Figure 1 shows the variation in optical density corresponding to the anti-gSG6-P1 IgG response from March to December 2005, from January to December 2006 and January 2007, with the LLINs being implemented in February 2006. The results obtained evidence:

large individual and inter-group variations of the IgG anti-gSG6-P1 response showing that each response is specific to the individual and the season (thus to the exposure period); - seasonal variations in median IgG anti-gSG6-P1 responses, despite the great diversity of individual responses. Indeed the maximum response is noted in May 2005 (peak) following the period of heavy rains (March-April). This maximum response is followed by a decrease in the IgG level in July-August-September (dry season), before increasing again in October (small rainy season). 20 - the introduction of the LLINs in February 2006 (before the heavy rains of March-April) is followed by a remarkable decrease in the IgG anti-Pl level (P <0.0001 in Mann-Whitney), so that in April (higher anopheline densities), the level of anti-P1 IgG response is very low or nil in most individuals, demonstrating the effectiveness of LLINs in reducing human-anopheles contact.

A comparison of the profile of individual IgG responses in January 2006 (the last month before the introduction of LLINs) and April 2006 (the month of sampling after LLINs were set up) was performed on the samples from 95 individuals during these two months. periods to better evaluate the impact of LLINs.

The results obtained are given in Figure 2.

It is found that the individual level of IgG falls significantly (P <0.0001) in April, 2 months after the introduction of LLINs, in almost all individuals.

Evolution of the number of answering machines during the study:

The number of responders was determined from a response threshold. This threshold was calculated in 14 French individuals not exposed to Anopheles stings according to the following formula:

Responder Threshold = Avg (ADO Individuals) + 3SD (ADO Individuals) 0.204: 30 This means that a subject is considered a responder if his ADO (anti-gSG6-Pl IgG response) is greater than 0.204. The percentages of answering machines per passage (or month) were calculated according to the following formula:% Responders = No. Responders / No. Individuals sampled during a passage. The number of losses, that is to say of absent individuals, per passage was also evaluated (see Table 1). Table 1: Percentage Loss Rates and Percentages of Responders Passages No. of No. of individuals of non-responding responders on whom responders of the samples were collected March-April 05 95 10 (9.52%) 72 23 (75, 78%) May 05 103 2 (1.9%) 81 22 (78, 64%) July 05 101 4 (3.81%) 77 24 (76, 23%) August 05 100 5 (4.76%) 69 (69%) 31 September 05 101 4 (3.81%) 78 23 (77.23%) October 05 99 6 (5.71%) 71 28 (71.71%) December 05 102 3 (2.86) 66 36 (64.71%) January 2006 100 5 (4.76%) 64 (64%) 36 Implementation LLINs (February 2006) April 06 100 5 (4.76%) 5 (5%) 95 June- Juilllet 94 11 16 78 06 (17.02%) (10.48%) August 06 101 4 (3.81%) 9 (8.91%) 92 October 06 96 9 (8.57%) 30 66 (31 , 25%) December 06 85 20 74 11 (19.05%) (87.06%) January 07 88 17 76 12 (86.36%) (16.19%) The highest loss rates are recorded in June-10 July (10.48%), December 2006 (19.05%) and January 2007 (16.19%), so after the implementation of LLINs. The results5 of the months concerned must therefore be analyzed with a certain degree of caution. On the other hand, the comparison between January and April 06 clearly shows the effectiveness of LLINs in reducing the level of anti-P1 IgG in the population. The results of the evolution of the percentages of responders are represented in FIG. 3:

Prior to LLIN implementation, the number of responders 10 varies with the season with a maximum in the rainy season of 78.64% in May and a minimum of 64.71% in December (beginning of the long rainy season). In the dry season the percentage of responders remains high with 76.23% in July, 69% in August and 77.23% in September. These results mean that the individuals involved are sufficiently stung to develop an antibody response during the dry season. The sharp decrease of the answering machines in April 2006, after implementation of the LLINs, attests the effectiveness of the LLINs on the reduction of the number of bites received and therefore probably on the transmission. The individual anti-P1 IgG responses in the responders throughout the study period were taken into account to better characterize them in terms of intensity.

The results are given in Figure 4.

This graph shows that the responses are stronger in the answering machines of May and October before the introduction of the LLINs and decrease in intensity, in number also, after the setting up of the LLINs. Thus, in addition to the responder percentage, it is important to consider the intensity of the responses in the case of LLIN exposure and efficacy evaluation by the IgG anti-gSG6-Pl.5 response. Comparison IgG anti-P1, medium parasite and anopheline densities

Average anopheline densities are expressed as an An number. gambiae / trap / night and parasitic densities in geometric mean.

Figure 5 shows the comparison between mean anopheline densities and anti-gSG6-Pl IgG response. In general, the densities of An. gambiae are low (maximum 4 yr. gambiae / trap / night) which indicates a low exposure context. Prior to LLIN placement, anopheles densities were positively associated with the IgG anti-gSG6-P1 response with a peak in May coinciding with the peak anti-P1 IgG response and low values in the dry season ( July-September). On the other hand, there is an opposite evolution between these two parameters after the implementation of the LLINs, in April-July 2006. Thus, it appears that the observed decrease in individual and collective IgG responses is not due to the absence of anopheles, but rather to the reduction of human-vector contact thanks to the physical and chemical protection that LLINs confer on populations using them. The comparison of IgG response and parasite densities is shown in Figure 6.

This figure reports a good association between anti-P1 IgG response and mean parasite densities before and after LLIN placement, in contrast to average anopheline densities. Indeed, the peak of parasitaemia (in May) before LLINs coincides with that of the IgG response and the decrease of parasite densities, after LLINs, with that of the marked IgG anti-P1 response. The fact that the IgG response curve is above and above that of the mean parasite densities, respectively before LLINs and after LLINs, demonstrates that the anti-P1 IgG response is a more sensitive tool than parasitaemia for evaluation. the effectiveness of LLINs.

2- Study of the influence of the factors of variation such as the age and the sex of the individuals on the IgG anti-P1 answer a) influence of the age on the anti-gSG6 P1 response The sample used was subdivided into 3 age groups: 0-6 years (n = 47), 7-14 years (n = 36) and +14 years (n = 22).

The results are shown in Figures 7 to 9: These figures show that the evolution of the antiSG6 P1 IgG response is the same regardless of the age range of the individual, both before and after use of the LLINs, and is similar to that of the total population (children + adults). These results indicate that the anti-gSG6-P1 IgG response is a: Anopheles sting exposure marker regardless of the age of the exposed LLIN efficacy indicator regardless of the age of the 25 subjects. using them.

A comparison was also made of the evolution of the IgG responses of these three age groups to check whether they are different in terms of intensity and thus levels or levels of IgG.

The results are shown in Figure 10. Examination of this Figure shows that differences in IgG anti-gSG6-P1 response intensity only appear at peak response levels (May and October) and January before use. LLINs and in June-July, 4-5 months after the LLINs are in place. In fact, individuals over the age of 14 (adolescents and adults) have a significantly higher anti-P1 IgG response than others during these periods. These older individuals would have received more bites than the older ones during these periods. In June-July, after LLINs, this effect is highly significant (P <0.0001), suggesting that older subjects tend to be more exposed to bites. For example, adults stay out longer for various reasons while children are forced to sleep under their LLINs. The anti-gSG6-P1 IgG response may therefore be an interesting indicator of the subjects' behavior towards the use of LLINs and other vector control strategies against malaria and other arthropod vector diseases.

Analyzes were also done to see if the implementation of LLINs is more effective in one of these age groups. For this purpose, the individual response profiles in each age group between January (last pass before LLINs) and April 2006 (1st passage after implementation LLINs) were evaluated.

The results obtained are shown in Figures 11A to 11C, with children up to 6 years of age (Fig. 11A), 7-14 years (Fig.11B) and above 14 years (Fig. 11C) .

These figures show that all subjects aged 0-6 years have an IgG level that decreases after LLINs and becomes very low or zero. In subjects aged 7-14 years, and up to 14 years, there are 2-3 individuals in whom the IgG level becomes higher despite the presence of LLINs. These results confirm the previous hypotheses on the behavior of individuals with regard to LLINs, their use and Anopheles.

The distribution of some parameters of central tendency (mean and median) and dispersion (minimum and maximum, therefore the extent of each distribution) in each age group and for each passage (month) was then studied.

The results are shown in Fig.12: the medians are represented by the horizontal bars, the averages by the "+" intra-boxes, the 0-6 years in red, the 7-14 years in blue and the +14 years in blacks.

This figure confirms that the intensity of the anti-P1 IgG response increases with age and is higher on average in the +14 years before LLINs, although the dispersion is less extensive in this group. These results suggest that individual responses are more heterogeneous in the first two groups, which therefore have greater variability. According to one aspect of great interest, the parameters of central tendency remain identical in the 3 groups in April 2006, 3 months after the setting up of the LLINs and the dispersion much more marked for the 7-14 years and +14 years thus confirming the less marked LLINs in two groups compared to 0-6 years.

Moreover in July, we begin to find the same evolution, but less marked, median and average than before LLINs suggesting a decline in efficiency of the latter.

Responders were also taken into account. The results given relate to the April 2006 answering machines to see their future in July and August (Fig 13) and to the responders in July 2006 to evaluate their IgG responses in April and August (Fig.14) of the same year. .

Of the 5 respondents in April 2006, 3 are 7-14 years old and 2 are +14 years old. It is noted that the individual response patterns in April vary in June-July and August, which confirms the specificity of the IgG response to exposure, and therefore to the season and character of the individual vis-à-vis the use of LLINs.

The same observations are valid for the month of June-July. Note that there are more responders in June-July, compared to August, and adults and children aged 7-14 (see Figures 15 and 16). These observations need to draw attention to the behavioral issues associated with implementing in place LLINs as an anti-vector strategy.

- Influence of sex on the anti-gSG6 response Pl The subdivision of the population studied by sex makes it possible to follow the evolution of the IgG response in 65 women and 40 men. Figure 17 shows the median evolution of the anti-gSG6-P1 IgG response in both groups throughout the duration of the study.

Examination of this figure shows that the evolution of the anti-gSG6-P1 IgG response is similar in both groups before and after LLINs and that it has no difference in IgG response between men and women. In addition, it appears that the anti-P1 IgG response similarly drops significantly in both groups after LLINs, suggesting that the gSG6-P1 peptide is a marker regardless of sex.

All of these results show that the peptide 1 of the Anopheles gambiae gSG6 protein is not only a low-exposure marker, but also an indicator for evaluating the efficacy of LLINs as a vector control tool. malaria. This peptide is a marker whatever the age and sex of individuals using LLINs. It could also provide insights into the behavior of these individuals with respect to the use of LLINs and exposure to vectors.10 References

1 - Remoue F., Cissé B., Ba F., Sokhna C., Hervé JP., Baker D. and Simondon F. Evaluation of antibody responses to Anopheles salivary antigens as a potential marker of risk of malaria? Trans. Roy. Soc. Too much. Med. Hyg. 2006; 100: 363-370. 2 - Cornelie S, Remoue F, Doucoure S, Ndiaye T, Sauvage FX, Baker D and Simondon F. An insight into immunogenic salivary proteins of Anopheles gambiae in African children. Malaria Journal. 2007; 6: 75. 3 - Poinsignon A., Cornelie S., Mestres-Simon M., Lanfrancotti A., Rossignol M., Baker D., Cisse B., Sokhna C., Arca B., Simondon F. and Remoue F. Novel peptide marker corresponding to salivary protein gSG6 possibly identified exposure to Anopheles bites. Plos ONE. 2008. 3: e2472. 4 - Poinsignon A, S. Cornelie, F. Remoue, P. Grebaut, D. Courtin, A. Garcia and F. Simondon. Human African Trypanosomiasis: Initial Screening of Immunogenic Salivary Proteins of Glossina Species. Am. J.

Too much. Med. Hyg.2007. 76: 327-33

5 - Drama PM., Poinsignon A., Besnard P., The Mire J., Dos-Santos MA. Sow CS, Doucoure S., N'Diaye A., Cornelie S., Foumane V., Toto JC., Sembene M., Baker D., Simondon F., Fortes F., Carnevale P. and Remoue F. Human anopheles gambiae saliva: a new immuno-epidemiological marker to evaluate the malaria benefit from anti-vector strategies based on Treated Nets Insecticide. Am. J. Too. Med. Hyg.2010. In press 6 - Poinsignon A., Cornelie S., Ba F., Baker D., Sow C., Rossignol M., Sokhna C., Cisse B., Simondon F. and Remoue F. Human IgG response specific to a salivary peptide, gSG6-P1, a new immunoepidemiological tool for anopheles cite. Malaria Journal. 2009. 8: 198

7 - Remoue F., E. Alix, S. Cornelie, C. Sokhna, B. Cisse, S. Doucoure, F. Mouchet, D. Boulanger and F. Simondon. Evaluation of IgE and IgG4 antibody responses to Aedes saliva in African children. Acta Tropica. 2007. 104: 108-115.

8 - Orlandi-Pradines E., Denis de Senneville L., Almeras L., Barbe S., Remoue F., Villard C., Cornelie S., Penhoat K., Bourgoin C., Fontenille D., Bonnet J., Pagés F., Laffite D., Baker D., Simondon F., Fusaï T., Rogier C. Immune response against malaria and arbovirus vectors in travelers in tropical Africa. Microb. Infect. 2007. 9 (12-13): 1454-6220

Claims (10)

CLAIMS1 - Use of immunogenic and specific salivary proteins of mosquitoes Anopheles and Aedes as biomarkers for evaluating the efficacy of vector control strategies in the control of malaria and arboviroses, these proteins being chosen from the group comprising proteins and peptides of sequences SEQ ID Nos. 1 to 30. 2 - Use of the human antibody response to immunogenic and specific Anopheles and Aedes mosquito-specific salivary proteins of sequences SEQ ID Nos. 1 to 30 according to claim 1 for evaluating the efficacy of vector control strategies in the control of malaria and arboviruses. 3 - Use according to claim 1 or 2, characterized in that the target Anopheles mosquitoes include Anopheles gambiae and Anopheles funestus complexes and Aedes mosquitoes include Aedes aegypti and Aedes albopictus. 4 - Use according to any one of claims 1 to 3, wherein the proteins or peptides meet the sequences SEQ ID No. 2, 7, 9, 10, 12 and 14. 5 - Method for evaluating the effectiveness of vector control strategies in the control of malaria and arboviroses, characterized in that it comprises the evaluation of antibody responses specific to immunogenic salivary proteins and / or peptides of Anopheles mosquitoes and Aedes as a criterion for efficacy of LAV, whatever the larvicidal or adulticidal form of LAV, these proteins and peptides being chosen from the group comprising the proteins or peptides corresponding to the sequences SEQ ID Nos. 1 to 30. 6 - Method according to claim 5, characterized in that the proteins or peptides correspond to the sequences SEQ ID Nos. 2, 7, 9, 10, 12 and 14. 7 - Method according to claim 5 or 6, characterized by the evaluation of the IgG antibody responses, and isotypes (IgG1, IgG2, IgG3, IgG4, IgM, IgE and IgA) against the proteins / salivary peptides defined in claim 1, in individuals living in areas where an LAV strategy is applied, before and after application of the LAV intervention protocol. 15 8- Method according to any one of claims 5 to 7, characterized in that the LAV strategy comprises the use of mosquito nets impregnated with insecticides and other forms of LAV by larvicides and adulticides. 9 - Method according to any one of claims 5 to 8, characterized in that it is applied to a population living in endemic zone. 25 10 - Method according to any one of claims 5 to 9, characterized in that it is applied to a population of travelers. 20