A NOVEL PASSIVE VACCINE FOR CANDIDA INFECTIONS

The present invention relates to a passive vaccine for use in the prophylaxis and treatment of Candida sp. infection.

Passive vaccination, the therapeutic or preventative use of antibodies or antigen-reactive antibody fragments, is a remarkably attractive means for the control of infectious diseases in an era of emerging and re-emerging infectious agents, increased antimicrobial resistance threat and paucity of new drugs and preventive vaccines ³ .

Passive vaccination is particularly attractive for the control of opportunistic infections which rely upon immune dysfunction to cause disease in a debilitated host. Absence or reduction of immune competence reduces the ability of the host to cooperate with antimicrobials and also makes it particularly hard to generate a protective vaccine.

Candida sp., including C. albicans, C. tropicalis and C. parapsilosis, are extracellular, opportunistic pathogens. In particular, Candida albicans is a major mucosal and systemic pathogen, and is particularly opportunistic in immunocompromised hosts, especially in neutropenic patients or those with AIDS. Importantly, it is also a frank pathogen in the setting of vaginal candidiasis and other mucocutaneous pathologies, causing disease in both normal and immunocompromised patients. It has been estimated that three quarters of all normal women in fertile age experience at least one attack of acute vaginal candidiasis, and more importantly, about 5% of them suffer from chronic recurrences thereafter.

Chronic vaginal candidiasis is barely curable with antimycotics. Indeed, there is increased concern that Candida may become refractory to antimycotics under repeated treatment ² . A remarkable amount of money is spent by these unfortunate women also on palliative, anecdotal cures with undemonstrated efficacy, such as herbal extracts.

Candida albicans has defined virulence traits and, most importantly, studies with experimental animal models of infection have recently demonstrated that antibodies of

the right specificity and isotypes can indeed protect against severe mucosal and systemic experimental infections. Some antibodies are in phase 1 clinical trials.

By using an experimental model of estrogen-dependent vaginal candidiasis in rats , we have previously shown that: /) the secretion of some Secreted Aspartyl Protease enzymes (SAP) by Candida is necessary for the mucosal disease; ii) SAP inhibitors cure the disease; Ui) that the use of active vaccines directed against SAP, administered intranasally or intravaginally, can lead to successful vaccination; iv) the protection is substantially mediated by antibodies against SAP.

In cutaneous infections, SAP enzymes are amply secreted by the fungus and degrade the keratinous layers of the skin. The same is true for vaginal infections where SAP enzymes are able to degrade both the keratinous layer overlying the epithelial cuboid tissue and protective soluble factors of innate or adaptive immunity such as the cytokines and antibodies

One of the ten members of SAP family, the S AP2 enzyme, is particularly relevant as a virulence factor, and so antibodies against SAP2 have been shown, in active vaccines, to be protective (Reference 5). Other members of the SAP family, including SAP 1 and SAP 3, show a high degree of homology with SAP2.

Table 1 below shows a sequence alignment of the conserved catalytic sites of several aspartyl proteases of both viral and eukaryotic organisms, providing the rationale for some form of shared susceptibility to protease inhibitors ⁶ . SAP2 of C. albicans is particularly similar at this site to HFV protease and the plasmepsins of P. falciparum.

Our previous studies, including WO 2006/097689, support the notion that antibodies may be particularly relevant for passive vaccination in mucosal candidiasis, in particular vaginal candidiasis ³ . However, as the SAP proteins are all themselves protease enzymes, attempts to use antibodies to neutralise SAP have been severely hampered by the fact that the protease enzymes degrade the antibodies before this can be achieved. Indeed, WO 2006/097689 makes no mention of the potentially devastating affects on Ab activity that the proteases can wreak.

Enzyme Sequence SEQ ID NO.

PepsinA AQDFTVVFDTGSSNLWVPSV 4

Renin PQTFKWFDTGSSNVWVPSS 5

Cathepsin D PQCFTWFDTGSSNLWVPSI 6

P. falciparum PM2 QQPFTFILDTGSANLWVPSV 7

C. albicans SAP2 KDKVSVSIDTGSYDLWVMSN 8

HIV-I pr GQLMEALIDTGADDTVLEEM 9

HTLV-I pr PKTIEALLDTGADMTVLPIA 10

Table 1 : Amino acids shown in bold and underlined show identity; Amino acids in bold show a high degree of homology; and Underlined Amino acids show some similarity.

However, we have surprisingly discovered a number of monoclonal antibodies, or their active domains, that are capable of recognising and neutralizing S AP2, or other highly conserved and similar SAP members, thus providing a beneficial therapeutic effect against Candida.

What is particularly surprising, however, is that it would appear that the antibodies are not degraded by the enzymatic activity of the SAP proteases, which would normally be expected to cleave most proteins, including antibodies. These antibodies are, therefore, useful therapeutics against mucosal, cutaneous and systemic infections by Candida sp., especially in a passive vaccine.

Thus, in a first aspect, the present invention provides a passive vaccine, for use in the treatment or prophylaxis of a Candida sp. infection, comprising an antibody, or a fragment thereof, capable of recognising at least one epitope from a Secreted Aspartyl Protease (SAP) enzyme from Candida sp., the antibody or fragment thereof being substantially resistant to degradation by the Secreted Aspartyl Proteases.

Antibodies and polynucleotides encoding them are also provided. The Antibody may be of any form, but monoclonal antibodies (mAb) are particularly preferred.

In a further aspect, the present invention also provides antibodies, or fragments thereof, capable of recognising an epitope from an SAP enzyme from Candida sp. the antibody or fragment thereof being substantially resistant to degradation by said enzyme.

A passive vaccine is provided, for use in the treatment or prophylaxis of a Candida sp. Infection. The vaccine comprises an antibody, or a fragment thereof, capable of recognising an epitope from a Secreted Aspartyl Protease (SAP) enzyme from Candida sp. The antibody, or fragment thereof, is substantially resistant to degradation by the Secreted Aspartyl Proteases.

Reference to one epitope or one SAPs may be understood to include one or more such epitopes or SAPs unless otherwise apparent.

When targeting protease enzymes that degrade proteins such as antibodies, one cannot be sure that, when generating an active vaccine, the antibodies generated will not be degraded by their protease targets.

However, the antibody discovered by the present inventor are substantially resistant to protease activity, in particular the aspartyl proteases, SAPl, SAP2 and SAP3, or other bacterial and host proteases possibly present in female vaginal fluids, such that they retain sufficient activity to allow immune clearance of the proteases. Immune clearance is thought to be through the stimulation of the complement pathways.

In particular, it is preferred that the antibodies are resistant to cleavage by the proteases, at least to extent that they retain at least 50% and more preferably at least 60%, more preferably at least 70%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, and most preferably at least 99% of their immune clearance ability in the presence of the protease, for instance under suitable conditions for the protease to cleave suitable target proteins. Such ability is preferably the ability of the antibody to bind to a protease and thereafter stimulate the complement pathway and lead to the ultimate destruction of the protease. This may be measured by modifying the proteases themselves, for instance at the active site, so that

their proteolytic function is reduced, thus allowing a comparison of the activity of the Antibodies in immune clearance of the albeit modified proteases.

Advantageously, the Ab according to the present invention binds to one or more SAPs as herein defined with a K _0S rate constant of between 5 xlO ^"1 and 1 xlO ^"7 s ^"1 .

More advantageously, the Ab according to the present invention binds to the SAP with a dissociation constant (Kd) of at least 100 microMolar to 1 picoMolar.

Candida infections of any species maybe treated using one or more Abs according to the present invention. Suitable Candida spp infections for treatment using the Abs of the present include any of those in the group consisting of the following: Candida ciferrii, Candida famata, Candida lambica, Candida lipolytica, Candida norvegensis, Candida rugosa, Candida viswanathii, Candida zeylanoides, Candida albicans, Candida tropicalis, Candida glabrata, Candida par apsilosis, Candida krusei, Candida lusitaniae, Candida kejyr, Candida guilliermondii and Candida dubliniensis.

Preferred Candida spp. for treatment according to the invention include any of those in the group consisting of the following: Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida krusei and Candida lusitaniae. Candida albicans is particularly preferred.

In a further aspect, the present invention provides an Ab according to the present invention for use in the prophylaxis and/or treatment of Candida spp. infection.

In a further aspect, the present invention provides an Ab according to the present invention for use in manufacture of a medicament for the prophylaxis and/or treatment of Candida spp. infection.

In a still further aspect the present invention provides a method for the prophylaxis and/or treatment of azole resistant Candida spp. infection in a patient by administering to a patient in need of such treatment an Ab according to the present invention wherein

the Candida is resistant to any one or more agent/s in the group consisting of the following: itraconazole, fluconazole and voriconazole, caspofungin, micafungin.

In yet a further aspect still, the present invention provides a method for the treatment of imidazole resistant Candida spp. infection in a patient by administering to a patient in need of such treatment an Ab according to the present invention wherein the Candida spp. infection is resistant to any one or more agents in the group consisting of the following: Clotrimazole, econazole, fenticonazole, sulconazole and tioconazole.

Also provided is a pharmaceutical composition comprising an Ab according to the present invention and a pharmaceutically acceptable carrier, diluent and/or excipient.

Preferably, the antibody has a Light Chain according to SEQ ID NO. 1, or a functional variant thereof.

Preferably, the antibody has a Heavy Chain according to SEQ ID NO. 2, or a functional variant thereof.

Preferably, the antibody has a Light Chain according to SEQ ID NO. 3, or a functional variant thereof.

It is particularly preferred that at least two of the three, and most preferably three, Complementarity Determining Regions (CDRs) of each chain are retained in any variant in order to maintain Ab specificity for the protease target.

Preferably, the Abs are humanized Abs. Preferably, the Abs are human variable domains or comprise human framework regions (FWs) and preferably one or more heterologous CDRs which bind specifically to one or more SAP proteins described herein. CDRs and framework regions are those regions of an immunoglobulin variable domain as defined in the Kabat database of Sequences of Proteins of Immunological Interest.

Thus, whilst it is envisaged that the CDRs are retained verbatim, and preferably within a similar framework, it is necessary for any variant to retain the ability to substantially resist degradation by the proteases. This will be readily apparent to the skilled person as any variant Abs can be readily assayed for their resistance to the proteases. An example is provided in the Examples section.

The CDRs of the Light Chain from NL2/2A8 are provided as SEQ ID NOS. 11, 12 and 13. Thus, it is particularly preferred that these are retained in any variant.

The CDRs of the Heavy Chain from NL2/2A8 are provided as SEQ ID NOS. 14, 15 and 16. Thus, it is particularly preferred that these are retained in any variant.

The CDRs of the Heavy Chain from NL2/9B9 are provided as SEQ ID NOS. 17, 18 and 19. Thus, it is also particularly preferred that these are retained in any variant.

It is also preferred that the antibody or functional variant thereof has 85% or more sequence identity to the respective SEQ NO., provided that at least two CDRs are retained.

Where reference to the retention of CDRs is made, it will be appreciated that minor modifications can be made to alter the specificity of the CDR for the epitope, for instance to increase or even decrease its specificity, but the latter is not particularly preferred. Such modification will have to be conservative in nature in order to retain the 3-d dimensions and hence the specificity of the CDR (i.e. the epitope binding pocket).

Suitable frameworks, also referred to as scaffolds, are known in the art and include human domain Abs (from Domantis), Camel/Llama V _H H (from Ablynx), Human TCRs (from Avidex) and so forth.

The advantage of altering the framework of the Ab is that the Ab can become increasingly humanised, although the need to retain resistance to protease degradation is still crucial.

Preferably, the vaccine comprises a mixture of antibodies NL2/ 2A8 and NL2/ 9B9, including fragments or functional variants thereof.

The functional variants have preferably 70% or greater sequence identity to one of said sequence ID numbers, preferably 80%, preferably 85%, more preferably 90%, more preferably 95%, more preferably 99% and most preferably 99.9% sequence identity to any one of sequence ID numbers 1 to 3.

Preferably, the antibody is a monoclonal antibody and, preferably, may also be a hybrid or chimeric antibody. Preferably, the antibody is Immunoglobulin type IgM or IgG.

It is preferred that the antibodies are those referred to herein as NL2/2A8, NL2/9B9, according to SEQ ID NOS. 1-2 and 3, respectively.

In a further aspect, the present invention also provides polynucleotide sequences, preferably DNA or mRNA, encoding the amino acid sequences of SEQ NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3.

It is particularly preferred that the antibodies are NL2/9B9 (SEQ ID NO. 3) or a functional variant thereof, or more preferably, NL2/2A8 (SEQ ID NOS. 1 and 2) or a functional variant thereof. These two antibodies show the greatest inhibition of SAP2 activity, as shown in Table 2, thereby providing the most therapeutically effective options for inclusion in a passive vaccine.

The passive vaccine may comprise only one type of the antibodies of the present invention, or may comprise a mixture of 2 or more antibodies. Particularly preferred is a vaccine comprising a mixture of the antibodies according to SEQ ID NOS. 1 - 2, and 3, including fragments or functional variants thereof.

It is a particular advantage of the A8 and B9 antibodies that they recognise different epitopes on each of the SAP sub-family of enzymes, comprising SAP 1, 2 and 3. In fact, these antibodies are shown in the example to be at least as effective as Pepstatin A, a known aspartyl protease inhibitor, and the known anti-Candidiasis drug, fluconazole.

Thus, it is particularly preferred that the passive vaccine has equivalent, i.e. statistically significant, or greater, efficacy compared to known treatments for Candidiasis, particularly Pepstatin A or fluconazole.

Passive vaccines are well known in the art, but the skilled person will understand that therapeutically effective and pharmaceutically acceptable titres of antibodies according to the present invention will be required. There are two broad categories of vaccines, active and passive. An active vaccine stimulates the host's immune system to produce specific antibodies or cellular immune responses or both, which would protect against or eliminate a disease. A passive vaccine is in general a preparation of antibodies that neutralizes a pathogen and may be administered before or around the time of known or potential exposure. Most references to the term "vaccine" per se are to active vaccines, which are the object of the vast majority of research and development activities in the field.

The passive vaccine may be administered intranasally or intravaginally. However, it is also envisaged that other modes of administration, such as oral, rectal, intravenous or parental administration could also be effective.

Alternatively, it is also envisaged that the antibodies may be expressed in the patient by means of gene therapy. Preferably, the vaccine will, therefore, comprise a suitable means for in situ expression of the antibodies, such as a suitable viral vector comprising polynucleotides encoding the antibody, preferably under the control of a suitable promoter, for instance.

It is also preferred that the passive vaccine may be administered in the form of a cream or ointment, the cream or ointment comprising a therapeutically effective and pharmaceutically acceptable titre of the above-mentioned antibodies according to the present invention. It is also preferred that the antibody can be part of a therapeutic cocktail which may include other drugs or other anti-Candida antibodies

Where reference to antibodies is made, it will be understand that this also includes functional variants thereof, as defined above.

hi a further aspect, the present invention also provides method of treatment or prophylaxis of a Candida sp. infection, comprising selecting a patient and administering an appropriate quantity of antibody according to the present invention, in the form of a passive vaccine, where the passive vaccine is as described above.

Furthermore, the invention also provides the use of an antibody or a functional variant thereof as described above, in the manufacture of a medicament for the treatment or prophylaxis of a Candida sp. infection.

The present invention is suitable for the treatment or prophylaxis of a range of Candidiasis infections, preferably mucosal or systemic Candidiasis. Vaginal Candidiasis is most particularly preferred.

Preferably, the Candida species is Candida albicans, but is also effective against other Candida species, particularly C. tropicalis and/or C. parapsilosis.

Preferably, the antibodies target and recognise epitopes on the SAP family of Candida proteases enzymes, most preferably the SAP 1-3 sub-family, including SAPl, SAP2 and SAP3. SAP2 is particularly preferred.

The epitopes recognised by the different antibodies of the invention may be the same, but are preferably different, such that each antibody preferably recognises a different epitope on SAP2, for instance. Due to the conserved nature of the protease family, it is preferred that each antibody recognises the same broadly conserved epitope on the different family members.

The fragment or functional variant of the antibody is capable of specifically binding to and recognising the epitope of the target enzyme. The fragment may be a particular portion of the Heavy or Light chains of the invention, and not necessarily just the variable domain, although this is preferred. Of course, restriction of the sequence may

alter the folding or reveal new protease sites, so the ability to resist degradation must still be assayed, although this I relatively simple, as discussed above.

Single domain antibodies are preferred, for instance using the methods those taught in WO 2006/097689 (Domantis Ltd), although resistance to degradation will need to be assessed.

The Ab may consist of only a single chain according to the present invention, or may simply comprise a single chain of the invention. Alternatively, the Ab may consist of more two, or possibly more, chains according to the present invention.

Without being bound by theory, we have discovered novel anti-SAP antibodies, which target SAP, being a critical virulence trait of the Candida fungus. We have shown that these monoclonal antibodies (mAbs) are capable of : ϊ) neutralizing SAP2 enzymatic activity; if) recognizing specific and distinct SAP2 epitopes, common with SAPl and SAP3 members of the family; and Hi) exerting a strong, fluconazole-comparable, therapeutic activity, particularly in vaginal candidiasis and also in strains of C. albicans resistanto to fluconazole The antibodies would appear to remain substantially active, despite the degradative activity of the SAP proteolytic enzymes.

The advantage of being able to deliver the antibodies in a passive vaccine is that active vaccines cannot be used in some circumstances, particularly in immunocompromised hosts in whom vaccines are unable to mount an effective immune response, even though it is in these hosts that opportunistic infections like Candida sp. axe most likely to occur. Furthermore, as mentioned above, one cannot be sure, when generating a protective immune response using an active vaccine, for instance, that the antibodies generated will not be degraded by their protease targets.

The invention will now be described by reference to the accompanying example, which is not intended to be limiting on the scope of the invention.

All references cited herein are hereby incorporated by reference, to the extent that they do not contravene the present teaching.

Example

Hybridoma generation and monoclonal antibody purification.

Two female Balb/c mice (Harlan) were immunized intrasplenically with 15 mg each of highly purified, recombinant, 6-his tailed SAP2 protein (Sandini et ah, submitted), suspended in 200 microliters of saline, followed by chronic boosting via subcutaneous injections at two weeks intervals of the same amount of protein suspended in Freund's adjuvant for three months. Four days after the last booster, the spleen was removed and fused with the X63.Ag8.653 cell line according to standard hybridoma protocols. Clones were screened through ELISA against the recombinant antigens, excluding all those reactive with another recombinant antigen (enolase), thus bona fide recognizing the histidine tail. Genuine SAP-binders were further cloned and selected for their capacity to inhibit SAP2 enzymatic activity using pepstatin A as positive control ⁶ . At the end of the procedure, two mAbs, named NL/2.A8 and NL/9.B9, were propagated as ascites and used for biochemical and functional testing. Both mAbs were determined to belong to IgM class by their SDS-PAGE profile and their reactivity in ELISA and Western blot assays with affinity isolated, alkaline phosphatase-conjugated goat anti- mouse M or G chain antibodies (Sigma).

Table 2 shows the ability of the NL/2.A8 and NL/9.B9 mAbs to neutralise the activity of SAP2. The data refers to a typical determination out of three performed with similar results. For methodology and definitions, see Ref.5, incorporated herein by reference.

Sample DeltaOD/ml/min % Inhibition

Buffer/BSA/SAP2 1.12 —

+ pepstatin A * 0.28 100

+ NL2/2A8 ** 0.44 82

+NL2/9B9 0.56 68

* taken conventionally as 100% inhibition in a spectrophotometric, BSA-degradation assay.

** with respect to pepstatin inhibition. Valid for all other inhibitors.

Table 2. S AP -2 Activity neutralization by mAbs

mAb Specificity and cross-reactivity within the SAP1-3 family members.

To investigate the nature of the epitopes recognised by mAbs, and their cross reactivity within the closely homologous SAP 1-3 members, full-length SAPl and SAP3 recombinant proteins were also generated. S AP2 itself was also cloned as two distinct partially overlapping fragments, named IB ₁ and 4Bj ₅ accounting for SAP2 amino acid sequences 57-158 and 117-203, respectively (see Reference 6). All recombinant products were tested together with an irrelevant 6-histidine tailed mAb (anti-enolase of C. albicans) as control.

Tables 3 and 4 show the antibody titres required for the NL/2.A8 and NL/9.B9 antibodies to recognise the SAPl, SAP3, SAP2 (full) and the two SAP2 fragment antigens.

Table 3 - Antibody reactivity with SAP 1-3 protein family and SAP-2 fragments*,** Reactivity with mAb NL2/2A8

Table 4 - Antibody reactivity with SAP 1-3 protein family and SAP-2 fragments*,** Reactivity with NL2/9B9

* Antigen used at a concentration of 200ng/ml. Antibodies were ascitic fluids. All ELISA positive reactivities were confirmed in Western Blot using polyvalent anti- mouse IgG.

There were no ELISA reactivities with any SAP or fragment using the irrelevant anti- enolase mAb.

As shown in Tables 3 and 4, both mAbs recognised SAPl and SAP3, in addition to SAP2, as expected, but differed from each other in their binding strength to the three SAP antigen proteins. Thus, both mAbs target an epitope common to the three SAP members. However, the epitope is not shared by the two mAbs as demonstrated by the observation that mAbNL2/2.A8 does not recognize either SAP2 fragment whereas mAb NL2/9.B9 strongly reacts with the IB ₁ N-terminus fragment of SAP2. Thus, each mAb binds to a different epitope, each epitope being common to all three SAP members. No reaction occurs of any SAP with the anti-enolase mAb.

Mab activity in the rat vaginal candidiasis model

The two mAbs described above, NL/2.A8 and NL/9.B, were thus assayed for their capacity to accelerate fungus clearance from the rat vaginal cavity challenged by a virulent vaginopathic Candida strain. The anti-enolase antibody (see above) was used a as negative control. The antimycotic drug fluconazole and the prototypal SAP inhibitor, pepstatin A, were used as positive controls. The model efficiency, significance and relevance for human disease has been discussed elsewhere ⁴

We found:

1. accelerated clearance during early treatment, i.e. three-six days from the intravaginal challenge; and

2. healing of infection ( <1 CFU/microlitre of vaginal fluid) on day 21 from challenge.

The differences were assessed by both parametrical (Student's t test) and non- parametrical ( Mann-Withney U test) statistics . The results are shown graphically in Figure 2.

All differences at each time-point (starting from day 1 post-challenge) between untreated/irrelevant mAb-treated (negative controls) and anti-SAP2 mAbs/pepstatin A and fluconazole-treated rats were statistically highly significant (P<0.01) by both parametrical and non -parametrical tests.

There was no statistically significant difference between pepstatin/fluconazole and anti- S AP2 mAbs-treated rats at any time-point. Five rats in duplicate experiments were used. Ascitic fluid (50 ul) was given intravaginally to each rat at hours -1, 24 and 48 from the infectious intravaginal challenge (10 ⁷ C. albicans cells , strain SA40, ISS collection). For other methodological details, see the De Bernardis et al. Aspartyl proteinases of Candida albicans and their role in pathogenicity. Med Mycol. 2001 Aug; 39(4); 303- 13. Review.

As seen in Figure 1 , both anti-SAP mAbs (2A8 and 9B9), but not the anti-enolase mAb (7C9), caused a highly significant acceleration of Candida clearance from the rat vagina, particular since the first day of treatment. In addition, the anti-SAP mAb-treated rats were nearly freed of Candida after the first week of infection, whereas all negative controls, including those treated with the irrelevant mAb, were still infected and could not clear the infection by the third week post-challenge. Importantly, the treatment with either anti-SAP mAb was as efficacious as the treatment with pepstatin A and a therapeutic fluconazole dosage (Figure 2).

Discussion

Both anti-SAP mAbs neutralize SAP2 enzymatic activity, though recognizing distinct epitopes of this enzyme, shared by at least other two members of the ten-member SAP family, i.e. SAPl and SAP3. It has to be stressed here that, firstly, SAP 1-3 sub-family expression is critically required for mucosal infection by Candida albicans, as demonstrated by gene knock-out experiments ⁹ and, secondly, that SAP virulence is explained by the capacity of these enzymes to hydrolyse a wide number of protein substrates, including immunoglobulins. Therefore, these mAbs are the first to neutralize SAP activity, rather than be degraded by the enzyme.

Furthermore, both anti-SAP mAbs exert a therapeutic effect in an estrogen-dependent rat vaginitis model, an effect which compares with the activity exerted by a prototypal inhibitor of aspartyl proteases (pepstatin A) and, more importantly, by fluconazole, a widely used and efficacious anticandidal drug.

References

1) Cassone,A & Polonelli,L. In : Novel vaccination strategies, S.H.Kaufmann Ed, Wiley, 2004, pp365-386.

2) Nyiriesy,A. & Sobel,J.D. Vulvovaginal candidiasis. Obstet Gynecol Clin North Am. 2003 ,30 :671-84.

3) Magliani et al. New immunotherapeutic strategies to control vaginal candidiasis. Trends MoI Med. 2002 ;8 : 121-6.

4) De Bernardis, F., Lorenzini, R. & Cassone, A. (1999). Rat model of Candida vaginal infection. In : Handbook of Animal Models of Infection (Oto Zak, E. & Sande, M. A., Eds), pp. 735-40. AcademicPress, New York, NY, USA.

5) De Bernardis,F. , Sullivan,P.A. and Cassone,A. :Aspartyl proteinases of Candida albicans and their role in pathogenicity. Med Mycol. 2001 39 :303-13.

6) Tacconelli,E. et al. Candidiasis and HIV -Protease inhibitors: the expected and the unexpected. Curr. Med. Chem. - Immun., Endoc. & Metab. Agents, 2004, 4, 67-84.

7) De Bernardis, F. et al. Local anticandidal immune responses in a rat model of vaginal infection by and protection against Candida albicans. Infect Immun. 2000 ;68 :3297- 304.

8) La Valle R. et al. Molecular cloning and expression of a 70-kilodalton heat shock protein of Candida albicans. Infect Immun. 1995;63 :4039-45.

9) De Bernardis et al. Evidence that members of the secretory aspartyl proteinase gene family, in particular SAP2, are virulence factors for Candida vaginitis. J Infect Dis. 1999 ,179:201-8.

Description of Sequences

SEQ ID NO. 1 (SEQ ID NOS 11 , 12 and 13 are underlined in order) Sequence of Light Chain for Ab from clone NL2/ 2A8:

γVMTOSPLSLPVSLGDOASISCRSSQSLVHSNGNTYLHWYLOKPGOSPKLLIYKV SNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSOSTHVPFTFGSGTKLEIK RADAAPTVSKYFVPEPγVI

SEQ ID NO. 2 (SEQ ID NOS 14, 15 and 16 are underlined in order) Sequence of Heavy Chain for Ab from clone NL2/ 2A8:

RKLSCAASGFTFSSFGMHWVRQAPEKGLEWVAYISSGSSTLHYADTVKGRFTIS RDNPKNTLFLOMKLPSLCYGLLGSRNLSHRLLSONDTPICLSTGOG

SEQ ID NO. 3 (SEQ ID NOS 17, 18 and 19 are underlined in order)

Sequence of Heavy Chain for Ab from clone NL2/ 9B9:

QLQOSGGGLVOPGGSRKLSCAASGFTFSSFGMHWVROAPEKGLEWVAYISSGS

STLHYADTVKGRFTISRDNPKNTLFLOMKLPSLCYGLLGSRNLSHRLLSONDTPI

CLSTGQGRIQA

Only the light chain of the Ab from clone NL2/ 9B9 has yet to be confirmed.

The 3 CDRs for each Ab are shown above, in order and underlined. Where variants of these sequences are provided, it is particularly preferred that the CDRs remain unchanged.