WO2011086535A1

WO2011086535A1 - Method for assessing the effectiveness of anti-vector control strategies in the control of malaria and arbovirosis

Info

Publication number: WO2011086535A1
Application number: PCT/IB2011/050210
Authority: WO
Inventors: Franck Jacques Louis-Marie Remoue; Anne Claire Poinsignon; Sylvie Caroline Cornelie; Papa Makhtar Drame; Souleymane Doucoure; François Jacques André MOUCHET; Marie Aude Bernadette Rossignol; Vincent Bernard Camille Corbel
Original assignee: Institut De Recherche Pour Le Developpement (Ird)
Priority date: 2010-01-18
Filing date: 2011-01-18
Publication date: 2011-07-21
Also published as: AP2012006415A0; FR2955393B1; FR2955393A1; EP2526418A1

Abstract

The invention relates to the use of immunogenic salivary proteins that are specific to the Anopheles and Aedes mosquitoes as biomarkers for assessing the effectiveness of anti-vector strategies in the control of malaria and arbovirosis, said proteins being selected from the group including proteins and peptides with the sequence SEQ ID No. 1 to 30.

Description

Method for evaluating the effectiveness of anti ^¬ Vector control strategies in the control of malaria and arboviruses.

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 by means of larvicides and adulticides, supports impregnated with insecticides, in particular mosquito nets, indoor sprays). 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 assess the degree of bites received by Man and thus the real effectiveness of LAV measurements tested or used.

These limits are particularly encountered in a context of low exposure to vectors, which is 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 allows to calculate the parasite prevalence and the intensity of infection which are tools used to evaluate the effectiveness of the LAV by comparing before / after establishment of the LAV. Major limitations relate to the need for long-term population monitoring (over a year, for example) before and after LAV implementation. Indeed, to affirm that the LAV did indeed reduce the prevalence or the intensity of Plasmodium infection, this evaluation must be carried out taking into account the season of transmission. These limits are particularly important in areas where 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 the detection of Plasmodium if antimalarial drugs are distributed to populations in the study area). In addition, the presence and intensity of infection of Plasmodium can vary very regularly from one day to another. As a result, it is very difficult to regularly monitor the populations of the area where LAV is assessed. This parasitological evaluation thus requires studies of longitudinal follow - up 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 costly logistical field resources.

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 answers specific antibodies to total or specific salivary extracts of the salivary proteins ^f Anopheles and Aedes represented relevant biomarkers to assess the exposure of human populations to the bites of the mosquitoes. More particularly, the research group to which the inventors belong has demonstrated that the total anti-saliva IgG antibody response of An, gambiae (1) and (2) and anti-peptide of the gSG6 protein of this vector (3) of Anopheles was a tool for assessing exposure to the malaria vector. In addition, the total anti-saliva 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 field, 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 evaluate the 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 present invention to provide a method for evaluating the efficacy of a 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 mosquitoes Anopheles and Aedes 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 sequences 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 Anopheles and Aedes mosquitoes of said sequences SEQ ID No. 1 to 30 to evaluate the effectiveness of vector control strategies in the control of malaria and arboviruses.

The invention also aims, according to another aspect, with a method of evaluating the efficacy of an LAV strategy in the control of malaria and arboviroses, characterized in that it comprises the evaluation of antibody responses specific to proteins and and / or immunogenic salivary peptides of mosquitoes Anopheles and Aedes as a criterion of 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 No. l to 30. The sequences SEQ ID No. 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: PI peptide of the gSG6 protein, hereinafter designated gSG6-Pl

EKVWVDRDNVYCGHLDCTRVATF

SEQ ID No. 3: P2 peptide of the gSG6 protein, hereinafter designated gSG6-P2

ATFKGERFCTLCDTRHFCECKETREPL

SEQ ID No. 4: P3 peptide 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 the gSG6 protein, hereinafter designated gSG6-P5

DRLGSCSKSMHDVLCDRIDQAFLEQ

SEQ ID NO: 7: gVAG protein

MAIWIACATLLLVVLSGVSAGGKYCSSDLCPRGGPHVGCNPPSSSGGPTCQGKQKARKVLLT PALQAYIMDEHNLNRSNIALGRIRPYPSAVKMPTLTWDPELASLADANARSCNYGHDRCRAT KKFPYAGQNIAITQFFGYRFTEKDLIHKFVSSWWSEYLDARPEHVRKYPSSYSGKPIGHFTQ IASDRSTKVGCSMWYWKDGQMDVYYFVCNYSFTNIMDRSVYQSGPTASQCKTGRNSKFPGLC NASEEPRS IMDP SEQ ID NO: 8 Protein 5 'nucleotidase

MALVRFA I ITVLCHLAIQDGAAKSFPHGEKAPFPLTLIHINDLHARFDETNQKSS CTNSK ECIAGIARVYHTIKQLKSEYKTKNPLYLNAGDNFQGTLWYNLLRWNVTAYFIKELPPDAMTL GNHEFDHSPKGLAPYLAELEKMKI PTVVANLEKNGEPALKDSKIAPQIVLKVGNRKVGVIGA LYDKTHLVAQTGMVTLTNS IEAVRKEAQELKKKNV I IVVLSHCGLDGDKQLAEEAGDLIDV IVGAHSHSLLLNKDAKVPYDTKYDTIEGDYPLVVKKSNNHTVLITQARSFGKYVGRLTVNFD CEGEVQSWEGYPIYMNNSVKQDEEVLRELEPWRAEVKRLGTQVIG EVFLDRESCRWCECT LGDLIADAYADQYTNSTVQPVAFVQAGNFRNPIEKGDITNGLAIEAAPYGSSVDMIKLSGAD LWSAIDHSFTLDDEFRYNTAQVSGMTVVADLSKKPYERVQS IDIVGANGAKTALKKDQIYYV AVPSYLADGKDGFAMLKKGTNRVTGPLDSDVLIEYVRKRQTIAKTMFQEKRVVIENHTNGTC SWDLDSQRYKPK

SEQ ID No. 9 SGl-like Protein 3

MAGQRHLIEQAWQYGAQLQHELMLTSMESDRVQRALVLHSMLVNASLAEMVKESYQTHGADG RMVVRMLKFVRLLPGADERVAVYKQLAELLKSNGQDGRFPAVIFSTDVRQLEDRYKPDHAQY EGKVVERWLAELQAGTFHEVVEFARDYPEYFARVEEPLYETLKQQWSAEGLDRMVSFPNALP VGVQRVRALRALLETLLQHQGEQNNDVYLIRLAHETGRVEATVGQADAAVRQALDDVKKLFE QFKYQRGFPDYEALYKLFKGL SEQ ID NO: 10: A protein 34 kDa No. l, with signal peptide (underlined)

MSPSNKILVLLLFPILLVSSHPI PAEDPAKQCNLSEDDLTKLKAAI SGAS SAKAANEDILPN TTLAACPMLKNFTEMLKTVATDMEVLKTQGVSNMEVQLLRESFEEKLNDLAKNKDIFERQAN QDTSKAEGEMVEKINKLQLEMAKLQEEIEEQTKQMYVDMIEYIFERLKMNDTEAIDSYAQIV MKTKMHELIMKLKTDRLVLWEMVKYVEGKKNKWVGRKVLNTILDQVNKLKLYKPEEVEIGKN SLVVVWCWKFNSETVYGTTDEDQKSFHLAKLFFPKEKGCKECANVKSRTMCNNDYPKVMVKA FG

SEQ ID No. 11: 34 kDa Protein No. 2, with signal peptide (underlined)

METSLPITVVFPIVLITGAQTKPTQGSCTLTDEDI SDIKSAVQKASKAAVNDIVLDPTLIDK CPMLEKITASLKSVATEIVQMRDSAISTDQVDQLKQNFEDQVNQIVKSRDIFEKQSGTQATK EHGEMLERMTAQVKVTELEQQIAKQTASMYEDMAELIFQRLQMNSTESVRSYTKHMMEEKLE ELMNKLE NYRIYLGALRFLNHMNDQELIGKVFDGILKRLGDMKADSDDVKENGRNLLVNLL CWTVNNDFLGKKYKERQVDLYRMALKFYPKTYEKAANEADVRSRQFCEENFPANLITWFAVS WNDRG

SEQ ID NO: 12 Protein 34 kDa # 1, with signal peptide (underlined)

Biomarker peptides are shown in italics.

MKTSLPIVVLLTAVISGVHP PrPXSC! RVSEEDLTTIRNAIQKASRASLDDVNLDEDLIAKC PLLK I ASLKSVASEIATLKDTGISEEQVDELKQSYEQQVNEIVKSROIFEKQSGGDVMKE QGAMINRMTELQVQVAQLQQQIGEQTSPMYDDMAELIFQRLAMNSTDSIRNYTAHMMEQKLH TLMTKLETNYRIFLGALRYLDHLGDQPLIDKVFDGILKRLDEMSLET KEPE GXYVLVNLL CWTVNNRFLTEKYRKKQLELFRIALKFYPKTG KEA EADIPGPQFCDANFPVNVITWFAVS RAAEGWGLRGTL

SEQ ID NO: 13 Protein 34 kDa No. 2, with signal peptide (underlined)

Biomarker peptides are shown in italics

MPVTYEFFILLAMSLLLVRSHPLPEEAITSDAAIKCTLSEEDLDSLKRAI SSAASAKSSOGII LSNETLTACPILANFTEMLKTVATDMEVLKTQGVSNAEVELLRESFEERLNEITKNKDIFER QAGQETSKTEGQMVEKINRLQLEMASLKEEIEQETiii YEDMAEYIFQRLKMNDTDAIDSYA QIMFKTKMHDLFMKLKTDRWVLWKMLNYVEQKKDKVIGKWVLNTVINQVTSL P XPDELEI GKDSLVNLLCWTSTAKTVYGAVQEDQKMFYLT

SEQ ID NO: 14: N-Term Peptide of the 34 kDa Protein

HPI PAEDPAKQCNLS

SEQ ID No. 15: 62 kDa Protein No. 1

MNPFTQVFCLIVGVLFFAFTATADEDCEI PLSELNKIEQTLTHIEKPIYTEEAESETSDSDE CAQMLRGIHLQLRRLTQKYKLMNKGYVKVEEFHKMAKEYEEQLSKLNGELEYFKVEARSGAD QKIKELKKDIKSLEQNVTKLHDRLKGITDELGKVSMELCSTYLESNQLDSAKEKVKDLPSKY VIELVDQFLSKNENNWLPVVDLSTAVPDLDDRGQVYK IYEFLLAKSRLEGEESLLLEAEIL KLNATFHPGSKITDDRKKELNDLLEKLRQNSEKTFEQWSQELDQS I SAVVFKNAIDRMFLT QIEKFGEYVTGRHDYYGVRNFLKLLGVSKNYHKIAAYNSLVEKKIGHALAVVMIDMLS Idia EIKHDPHVPDRIERMYNSTINSLPDSLKGI I PCIKSVKIRNEGTNRCIYAINQ IQVQNPNF K IVLGQYKRVTSTMSACTSFRLELSSNKPYIKI ITSEGNSLT INSAVPGETGFSRVGAPY SNKPNMKLDHTGDWVLDANFVNNS IKIQSDFNAYQ SKS IDHLVVRNDGDVAVVRYGLSGMR SGGI PDNHAEWKF

SEQ ID No. 16: 62 kDa Protein No. 2

A MNPFTQVFRLTVGVFFFAF ADEVCEI PLSELNKIEQTLTHIEKPIYTEEAESE SDSDE CAQMLRGIHLQLRRLTQKYKLMNKGYVKVEEFHKMAKEYEEQLSKLNGELEHFKVEARSGAD QKIKELKKDIQALEQNVTKLHDRLKGITDELGKVSMELCSTYLESNQLDSAKEKVKDLPSKY VIELVDQFLSKNENNWLPVVDLS AVPDLDDRGQVYK IYEFLLAKSRLEGEESLLLEAEIL KLNA FHPGSKI DDRKRELNVLLTKLSQFSEKTLEQWTQELNQS I SAVVFKNAIDRMFLT QIEKFGEYVTGRHDYYGVRNFLKLLGVSKNYHKIAAYNSLVEKKIGHALAVVMIDMLS Idia EIKHDPHVPDRIERMYNS INSLPDSLKGI PCIKSVKIRNEGTNRCIYAINQ IQVQNPNF I K S IVLGQYKRV MSAC SFRLELSSNKPYIKI ITSEGNSLT INSAVPGE GFSRVGAPY SNKPNMKLDHTGDWVLDANFVNNS IKIQSDFNAYQ SKS IDHLVVRNDGDVAVVRYGLSGMR SGGI PDNHAEWKLNALK

SEQ ID No. 17: 62 kDa Protein No. 1

MNSVNRYVLCIALIALFVASF AEENCEI SANELGKIEQTLTHINQPIYTGDDESEVSDSD QCAQMLRGIHFQLRRLTQKYKLMNKGYVKAEEFAKMARDYEDQLSVLKNDLEQSKIGADSSA KQKMQELKKDIATLEQNVNTLHKDLEGITDELGKVRMDLCLTYMESNQLSNAQDKVKTLAPK YLMELVEQFLNKSEKNWLPVVDLSVAI PDLDDRGQVYKTVHEFLKTKNRDGGEDS ILLEAEV LKMNA FHPGSKI EDRKKEIQDLLEKLSL S KIFDQWTQDLAKLENSAVYKNS IDRMFLT QMEKFGERVMAKDDYYSLRNFLKLLVVSTNYYKIAAYRKLIQEKIGHTLAVLMFDMMSMERT ELQYDPHVPDEVVRMYDES ITALPDSLKNIRSCLKLVKIYNHVTNQCILATNEVEDVNNSNP KFKSNVLGRRKLVKTASNDC PFRLEPSADKAS IRI ITPKGDALTNINS IQPGLSWFNRVGA PYTNNHNMKLDYSADWILDANYANDS IKIESEFNAYQTMKSVDHLMVTNVGKVPHVVVAQYG LKGMEYAGAGMKDAEWKFKCDN

SEQ ID NO: 18 Protein 62 kDa # 2

MKVYICQVIFSFLAVSVFCEENCNI PESELSKIDHVLRHMEKPIYSEEQFASDNEECTNLLN GIHAQLRRLTQRYKLMNKGYVKVEEYQRMADNYEKQLKTLNDELVELQQHTSEKASATIAKL KEDIKKLDEEVGTLHEKLKGIKQDFEKVKRDLCVTYLNSNQMSKAKAKLKEMASTYLIEIVQ QQLNKSNANIMPMLEFSAAI PDLDDMGEAYKEIYKFLEEQKRLEGEDSVLLEA VLKMNASL KEGSNITDERRTQIEGLLKDLATKSTIVFSTWTKELKKINDAVVIKNALDHMFVSQMKVFGA LVGD SDFGS IRNFVKL IVCNNYYKVAAYKELIDRKIGNALG IMFDLLTLEVNEMKFDPH VPDEI PKLFEATLSSLPNSLTELRTCLGKVQIYNKKTNKCVVATGNDFDVHKDKLGDFYRVV VADYGCTSFRLEASGDKASVRIVTPSGNPMSNVNLHLEGNSLHNYVATPKSNKPDRTPSSSD EWILDANYNNDTIKIESQFSDYKTKKTEVDHLLVRDINHLPHVLVARYGFMGLKNSDAKDTI EWNLKCGS

SEQ ID No. 19: 62 kDa Protein No. 3

MKWSVYIALLVFAFL SPVFSEENC I PESELSKIDDVLRHMEKPIYSEDHY SNNEECTNL LNGIHAQLRRLTQRYKLMNKGYVKVEEYKRMAEDYENQLKTLNAELLELQEHTSDKANAAIA KLKEDIKKLDEDVDTLHNKLKGIKQDFEKVKRDLCLTYLNSNQMSNAKAKVKEMASTYLIEI IQQRLNTKYA I I PMLDFS AI PDLDDRGEAYKEIYKFIE HERLDGEDAVLLEASLLKMNA TLKEGS I DERRTEIEKMLKELAEKSAVVFKTWS ELKGIED I IKYALDHLFVNQMKVFG GIVGDTFEFAPIRHLLKLLVVCNNYYKVAAYKELIDRKIGNVLGTIMFDLTTLEANEMSFDL HVPDEI PKLFNATLGSLPNSLTQLLPCLNKVHVYNAKTNMCIVAPEDRFDVQQEKL DFHRV VLAKYGCTAFRLESSPNKASVKFVKPSGNALSSINLQLENDQWHSHVGTPTANKPDRKPSSS DEWILDANYVNDTVKIQSEFNEYKASQAEVDHLLVMDVKYLPHVVVGRYGVRGLKRSSAKDT IEWYLKCAS

SEQ ID NO: 20 Protein D7 Long Form No. 1

MKLPLLLAIVTTFSVVASTGPFDPEEMLFTFTRCMEDNLEDGPNRLPMLAKWKEWINEPVDS PATQCFGKCVLVRTGLYDPVAQKFDASVIQEQFKAYPSLGEKSKVEAYANAVQQLPSTNNDC AAVFKAYDPVHKAHKDTSKNLFHGNKELTKGLYEKLGKDIRQKKQSYFEFCENKYYPAGSDK RQQLCKIRQYTVLDDALFKEHTDCVMKGIRYITKNNELDAEEVKRDFMQVNKDTKALEKVLN DCKSKEPSNAGEKSWHYYKCLVESSVKDDFKEAFDYREVRSQIYAFNLPKKQVYSKPAVQSQ VMEIDGKQCPQ

SEQ ID NO: 21 Protein D7 Long Form No. 1

MNILLVLAVV I SSLALVTAKGPFDPEEMHFIFTRCMEDNLKDGPDRVKTLLKWKEWVTEPK DDPATHCFAKCVLEMSGLYDAASGKFDASVIEAQHKAYPNSEDKGKVDAFVKAVQALPPTKN DCTAVFRAFGPVHMAHKA S INLFHDNKALTKGIYEKLGKDIRQRKQSYFEFCENKHYPVGS PKRSDLCKIRQYVVLDDAQFKQHTDCIMKGLRYITKDNILNCDEIKRDFKQVNKDTGALEKV LNTCKAKEPRDVKEKSWHYYKCLVESSVANDFKEAFDYREVRSQNYGYHLMQKQPYNKPAVQ AQVSEVDGKQCPS SEQ ID No. 22: 30 kDa Protein (allergen) No. 1

MKPLVKLFLLFCLVGIVLSRPMPEDEEPVAEGGDDDASGESEGEEETTDDAGGDGGEEENEG EEHAGDKDAGGEDTGKEENTGHDDAGEEDAGEEDAGEEDAGEEDAGEEDAEKEEGEKEDAGD DAGSDDGEEDSTGGDEGEDNAEDSKGSEKNDPADTYRQVVALLDKDTKVDHIQSEYLRSALN NDLQSEVRVPVVEAIGRIGDYSKIQGCFKSMGKDVKKVI SEEEKKFKSCMSKKKSEYQCSED SFAAAKSKLSPITSKIKSCVSSKR

SEQ ID NO: 23 Protein 30 kDa (allergen) n ° 2

MKPLVKLFLLFCLVGIVLSRPMPEDEEPVAEGGDEETTDDAGGDGGEEENEGEEHAGDEDAG GEDTGKEENTGHEDAGEEDAGEEDAGEEDAGEEDAEKEEGEKEDAGDDAGSDDGEEDSTGGD EGEANAEDSKGSEKNDPADTYRQVVALLDKDTKVDHIQSEYLRSALNNDLQSEVRVPVVEAI GRIGDYSKIQGCFKSMGKDVKKVI SEEEKKFKSCMSKKKSEYQCSEDSFAAAKSKLSPI IKSCVSSKGR SK SEQ ID NO: 24: A protein 30 kDa allergen # 3

MKYLLTFLMALSLVNLMLTRPTPEDDGGTSEEPQTQETTGSDEKNGASEEPNADDASKPDDV EEKGDDDTAKKEDDGESKDGEGSEKSDKEKGEPKNDPRETYNKVIEQLDQIKVDNVEDGHER SELAADIQRYLRNPIVDVIGSAGDFSKIAKCFKSMVGDAKKAIEEDVKGFKECTAKKDSNAY QCSQDRSTVQDKIAKMSSKIASCVASNRS

SEQ ID NO: 25 Protein 30 kDa, allergen no.

MQLFPI SLVVSCLVFSFVLSLPLPEEEAS SEE EA EAESAAADE ATGADDAESGSSE KNNEGGETSAPEAKEEGSGKDDRLDTYNKVNELLDKGTGVNNVENGFLRAFLDNNLQSHLRV PVIDAIGLVGDFSKIAGCFTSMGADVKKIVDDKLKSFTTCKSGKEAGDAKCSEDGSSVGGQQ DQITSKIKECVSSKGKGK

SEQ ID N ° 26 Protein 30 kDa, allergen n ° 2

MRSLLVLFAALCLVGFVLAKPI PEDDTEA EEPKAEDAAEQ AEDAGGSDGEKNDDEKPS DDAASNDQPKDVEGGEDGKESPKTEGEAKNDPQETYNKVIELLDQIKVDNVENGYERSELTS DLQTYLRNPIVDAITVGDFSKIADCFKSMGDEAKKAIEEELKAFKECTDKKESDAYQCSKNH STVQGKIAKISSTINSCVVSKRA SEQ ID NO: 27 Apyrase protein / 5 'nucleotidase No. 1

MARLSFVPLVI SFLSCTTALSEKSLFPLS I IHVNDIHARFEETNYVGTVCKPHEGQVCIGGY ARTVTVVKHLLRIRQNPLYLNAGDNFQGTLWYNMFRWNVTSTFLNMLPADAMTLGNHEFDNG VDGVI DCFP ASSPVLLCNVDASEVPEFVKYEKS IVIERAGRKIGI IGVILKN IVR SNPGK LRFLDEASSVKSEAQILKNQGIDI IVVLSHCGVDVDRRIATEGGEHVDI IVGGHSHTFLYSG NNTGI PGTVQGSYPTVVTHASGHKVLIVHAAAYTKLVGDIVLYFDEQGI IQRWEGNPHYLDP EVVPDPEVHKALLPWKLAVDELGNRI IGT I DLPRLGCNYRECALGNLI SDAFVDSSAYMA EPGHWSYAS IGLTNTGGIRVGLSKGDI SYKSLIEVI PFENTVDFIELRGDHLLRVLEHGTAK SNIQI SGLRVVYNMTEPYGNRVKSVQARCHGCSEPRFDPLDPFKYYRVAVNSHMAGGGGGFT MFEEFGRNKVVGDVEINALVRFVQKRSPLRYEVEGRVTVLN

SEQ ID NO: 28 Apyrase protein / 5 'nucleotidase # 2

MARLSFVLLVLLFLTYMTVIAEKPLFPLS I IHVNDIHARFEETNYVGTVCKPHEGQVCIGGY ARTVTVVKHLLRIRQNPLYLNAGDNFQGTLWYNMFRWNVTSTFLNMLPADAMTLGNHEFDNG VDGVI PFLDTVSSPVLLCNVDASEEPEFVKYEKS IVIERAGRKIGI IGVILKN IVR SNPGK LRFLDEASSVKSEAHILKDQGIDI IVVLSHCGVDVDRRIATEGGEHVDI IVGGHSHTFLYSG NNTGI PGTVQGSYPIVVTHASGHKVLIVQAAAYTKLVGDIVLYFDEQGI IQRWEGNPHYLDP EVVPDPEVHKALLPWKLAVDELGNRI IGTTMTGLPRLNCNYRECALGNLI SDAFVDSSAYMA EPGHWSYAS IGLTNTGGIRVGLSKGDI SYKSLIEVI PFENTVDFIELRGDHLLRVLEHGTAK SNIQI SGLRVVYNMTEPYGNRVKSVQARCHDCSEPRFDPLDPFKYYRVAVNSHMAGGGGGFT MFEEFGRNKVVGDVEINALVRFVQKRSPLRYEVEGRVTVLN

SEQ ID No. 29: apyrase protein / 5 'nucleotidase # 3

MLRTSSIVFLTCCLTFLIEGSSFKLKI IHFNDIHARFDEVTNSSSPCSGNGETCVAGIARLV TTIEKLRKQNENHLVLNAGDVFQGTIWYTLLKWNVSQQFMNMVKADAMTLGNHEFDDSFPVL I PFLENTKNVTPVVVSNLVFPKQLSRDVTKFRSLIKEDPLVLTVGGQS IGI IGVIFDETDKI GNADPLKFKSS IETVRIAAKQLKSKGVNI I IVLSHCGVFDDKKIAEQAGEDIDI IVGGHTHT LLYNGDPPSKHAALDKYPIVVETGNNHKVLIVQAFCHGHYVGNIDLTFDDEGEITAFEGQPI YQENRIEKNALVEARVRELRKDVEVKSLVKVGESKLELSNDCRLKDCTFGSVLADAYVWHFR SRSNAPMIAMIHPGNFRI SLAAGVITRGQILTALPFNSNANRVTVLGSTIKKAIEFGTS INP RRCSFNALQTAGIKIDVDYGKPVGNRTVILLKTGGKYKRLVESKKYDILVNSYVFKGGDGFD MFKHLAVKGRAPFDAELLEQYIVARKGIQKSGLLQSRMNVSHVEKALSEVKSCKQR SEQ ID NO: 30 Apyrase protein / 5 'nucleotidase No. 1

MAGKPGIQLFVIFILLSSFAAVVWTTDNMPADKDVSKLFPLTLIHINDLHARFDETNMKSNA CTAKDQCIAGIARVYQKIQDLLKEYKSKNAIYLNAGDNFQGTLWYNLLRWQVTADFI KLKP TAMTLGNHEFDH PKGLAPYLAELDKAGI PTLVANLVMNDDPDLKSSKIQKS IKVTVGGK I IM IGVLYDKTHEIAQTGKVTLSNAVETVKREAAALKKDKVDI IVVLSHCSYDEDKKIAKEAG QDIDVIVGAHSHSFLYSKESNKPYDQKDKIEGPYP IVESNNKRKI PIVQAKSFGKYVGRLT LYFDNEGEVKHWEGYPEFIDNKVKQDPKILEALI PWRKKVQEIGS KVGET IELDRDSCRD KECTLGVLYADAFADHYTNSSFRPFAI IQAGNFRNPIKVGKI NGDI IEAAPFGS ADLIRL KGDSLWAVAEHSFALDDENRTNCLQVSGLRIVIDPSKKIGSRVVKIDVMDNRNPKSEDLKPL DKNAEYFIALPSYLADGKDGFSAMKKATARWTGPLDSDVFKSYVEKIKKVDKLKWGRVIVCK AGSPCT

These immunogenic salivary proteins and / or peptides derived from these immunogenic proteins have been identified as immunogenic proteins and peptides and specific to Anopheles or Aedes and as biomarkers of exposure to pitting of the vector.

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 the LAV strategies against Anopheles mosquitoes and Aedes 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 the specific antibody responses to these salivary proteins as a criterion for efficacy of LAV, irrespective of the larval or adulticidal form of LAV.

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

The sequences SEQ ID No. 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, if appropriate, Anopheles funestus,

SEQ ID NO: 1, 11, 27-30, from Ae. aegypti

-SEQ ID Nos. 12, 13, 15, 16, 25, 26 and 30, of Ae. albopictus.

Surprisingly, the PI peptide 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, in accordance with 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 anti-saliva IgG response thus represents a new immunoepidemiological and individual indicator to evaluate the efficacy of anti-vector strategies against malaria, such as the use of insecticide-treated mosquito nets and other forms of insecticide-treated mosquito nets. of LAV 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 in 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, 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:

- Figure 1, the evolution of the IgG anti-gSG6-P1 response during the follow-up period (before and after establishment of long-lasting insecticidal nets LLIN abbreviated);

Figure 2, the impact of LLINs on the decline of anti-gSG6-Pl IgG responses;

- Figure 3, the number of answering machines according to the months before and after LLINs;

FIG. 4, the profile of the anti-Pl IgG responses of the responders;

Figure 5, the comparative evolution of the mean densities of An, gambiae and the median Pl-IgG response;

FIG. 6, the comparative evolution of the mean parasite densities and the median Pl-IgG response;

- Figures 7-9, the evolution of IgG anti-gSG6 PI response in 0-6 years, in 7-14 years, and in +14 years;

- Figure 10, the comparative evolution of the IgG anti-gSG6-P1 response in different age groups;

Figures 11A-11C, the profiles of individual anti-PI 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, April respondent profiles in July and August 2006 (Fig.13) and July in April and August (Fig 14);

- Figures 14 and 16, the age distribution of answering machines from June to July 2006 and the number of persons according to the age of answering machines from June to July;

17, the comparative evolution of the IgG anti-gSG6 -PI response by sex before and after LLINs. Example 1: Validation of the P1 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 are reported below, before and after using LLINs.

The most important malaria vector in this region is An. Gambiae s .1.

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

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

The results show: significant individual variations within and between groups ^¬ anti-gSG6-Pl IgG response showing that each answer is specific to the individual and season (thus the exposure period);

- seasonal variations of IgG anti-gSG6-P median 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). - 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 April (higher anopheline densities) the level of IgG anti-P1 response is very low or nil in most individuals, reflecting 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 implementation of LLINs) and April 2006 ⁽¹ month sampling after implementation of LLINs) was performed on samples from 95 individuals during these two 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 decreased 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 for 14 French individuals not exposed to Anopheles bites according to the following formula:

Responder threshold = Avg (ADO individuals) + 3SD (ADO individuals) = 0.204:

This means that a subject is considered a responder if his ADO (IgG anti-gSG6-P1 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: Loss Rates and Percentages of Responders

The highest loss rates are noted in June-July (10.48%), December 2006 (19.05%) and January 2007 (16.19%), so after the implementation of LLINs. The results 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 the LLINs in reducing the level of anti-Pl IgG in the population.

The results of the evolution of the percentages of responders are represented in FIG. 3:

Before LLINs were set up, the number of responders varied with the season, with a peak 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.

Individual responses of IgG anti-P1 in 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 answering machines in May and October before the implementation of LLINs and decrease in intensity, also in number, after the implementation of 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 assessment by the IgG anti-gSG6-Pl response. Anti-Pl IgG comparison, mean parasite and anopheline densities

Average anopheline densities are expressed in numbers of years. gambiae / trap / night and parasitic densities in geometric mean.

Figure 5 shows the comparison between mean anopheline densities and IgG anti-gSG6-Pl response.

In general, the densities of An, gambiae are low (maximum 4 An. Gambiae / trap / night) which indicates a context of low exposure. Before LLIN placement, anopheles densities are positively associated with the IgG anti-gSG6-P1 response with a maximum in May coinciding with peak anti-P1 IgG response and low values during 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 of 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-Pl 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 parasitic densities, after LLINs, at that of marked anti-Pl IgG 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-Pl IgG response is a more sensitive tool than parasitaemia for evaluation. the effectiveness of LLINs.

2- Study of the influence of the factors of variations such as the age and the sex of the individuals on the IgG anti-Pl answer a) influence of the age on the anti-gSG6 PI 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 IgG anti-gSG6 PI response is the same regardless of the age group of the individual, before and after using LLINs, and is similar to that of the total population (children + adults). These results indicate that the anti-gSG6-Pl IgG response is:

exposure marker to bite Anopheles irrespective of the age of the exposed subject

indicator of effectiveness of LLINs regardless of the age of the subjects using them.

A comparison was also made on the evolution of the IgG responses of these three age groups to check if 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-Pl response intensity appear only peak response levels (May and October) and January before LLINs are used and June-July, 4-5 months after LLINs are in place. In fact, individuals over the age of 14 (adolescents and adults) have a significantly higher anti-Pl 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 pass 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.llB) and above 14 years (Fig. 11C) . These figures show that all subjects aged 0-6 years have an IgG level which decreases after LLINs and becomes very low or even 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 about 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-Pi 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 for see their fate in July and August (Fig. 13) and 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 PI response

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 IgG anti-gSG6-P1 response is similar in both groups before and after LLINs and that there is 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 these results show that the peptide 1 of the gSG6 protein of Anopheles gambiae is not only a marker of low exposure, but also an indicator to evaluate the effectiveness 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 vector exposure.

Bibliographical references

1 - Remoue F., Cissé B., Ba F., Sokhna C., Hervé JP., Baker D. and Simondon F. Evaluation of anabolic 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., Boulanger 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. Med. Hyg.2007. 76: 327-33

5 - PM Drama., 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., Boulanger D., Simondon F., Fortes F., Carnevale P. and Remoue F. Antibiotic antibody response to Anopheles gambiae saliva: a new immuno-epidemiological marker to evaluate malaria efficacy use of Treated Nets Insecticide. Am. J. Too. Med. Hyg.2010. In nears 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 specifies a salivary peptide, gSG6- Pl, a new immuno-epidemiological 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., Aimeras 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., Fusai T., Rogier C. Immune response against malaria and arbovirus vectors in travelers in tropical Africa. Microb. Infect. 2007. 9 (12-13): 1454-62

Claims

1 - 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 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 salivary proteins of Anopheles and Aedes mosquitoes of sequences SEQ ID No. 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 the immunogenic salivary proteins and / or peptides of Anopheles mosquitoes and Aedes as a criterion for the 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 NO: 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.

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. 10 - Method according to any one of claims 5 to 9, characterized in that it is applied to a population of travelers.