Tissue reaction in guinea pigs after intramuscular application of living vaccine against trichophytosis.
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Vaccines of killed whole cells of Coccidioides immitis have been injected intramuscularly in 97 human subjects to determine the safety and tolerable dose of vaccine and certain immunologic responses. Seventy-eight individuals received the whole killed spherule preparation; 18 received killed mycelial vaccine. The maximum tolerable dose of spherule vaccine was approximately 5 mg. Systemic effects were slight though in two prior coccidioidin reactors a moderate febrile illness and sbustantial antibody production followed vaccination with killed spherules. The vaccines irregularly induced antibody or delayed hypersensitivity to coccidioidin or spherule antigen. Greater antigenic mass perhaps as a soluble preparation may be needed for consistent immunologic stimulation though the spherule vaccine may be protective despite absence of detectable antibody or sensitivity to coccidioidin.
In the Damage-response framework of microbial pathogenesis, infectious diseases are one outcome of a host-microorganism interaction in a susceptible host. In cryptococcal disease, damage to the host is caused by Cryptococcus neoformans virulence determinants, the nature of the host response, or both. Further, the disease may be acute or reactivated from a latent state. Hence, a vaccine for C. neoformans would need to prevent disease resulting from either acute or reactivated infection. The evidence to support the development of a vaccine for C. neoformans that induces antibody-mediated immunity is discussed herein.
Subcutaneous vaccination with recombinant antigen 2/PRA (rAg2/PRA) protected BALB/c mice against intranasal infection with Coccidioides immitis. Subcutaneously vaccinated C57BL/6 mice and intranasally vaccinated BALB/c mice were protected against larger numbers of infecting spores. Weight loss correlated with lethality, but histologic appearance did not. These studies support rAg2/PRA vaccination to prevent coccidioidomycosis.
Converse, John L. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.), Steven P. Pakes, Ernest M. Snyder, and Merida W. Castleberry. Experimental primary cutaneous coccidioidomycosis in the monkey. J. Bacteriol. 87:81-85. 1964.-Primary cutaneous coccidioidomycosis was studied in the monkey (Macaca mulatta) to find a suitable strain of Coccidioides immitis for use as a viable vaccine. Intradermal inoculation (medial surface of the right forearm) produced more severe vaccination reactions (draining vaccination site and axillary lymph node hypertrophy) than did subcutaneous injection. A subcutaneous vaccine dose of 10 arthrospores resulted in less reaction than a 100-spore dose. Moreover, dissemination beyond the regional lymph nodes did not occur after injection of ten spores of even the most virulent strains of C. immitis. Two of the five strains tested (Silveira, M-11, D-76, Cash, and a colonial isolate from Cash designated CW1) exhibited very mild vaccination reactions and appeared to have been cleared from the tissues upon necropsy at 10 months postvaccination. These two strains (Cash and CW1) appear promising for further immunological studies with a viable vaccine.
An understanding of the molecular bases of pathogenicity in Blastomyces dermatitidis and related systemic dimorphic fungi has been limited until recent years. Yeast cells of B. dermatitidis display an adhesion promoting protein termed WI-1. Recent studies entailing homologous gene targeting and mutation of WI-1 have provided null mutants at this locus and demonstrated the crucial role of the WI-1 adhesin in pathogenesis of blastomycosis. Ongoing studies are pointing to a link between phase-specific expression of WI-1 and the observation that transition to yeast cells is essential for the acquisition of pathogenicity by B. dermatitidis. Recombinant attenuated yeast that lack WI-1 are serving as invaluable tools for induction of vaccine resistance and are pointing to new insights about adaptive immunity to B. dermatitidis.
The possibility of the specific prophylaxis of the immunosuppressing effect of the products of microbiological synthesis (MBS) on laboratory animals by the preliminary immunization of the animals with the vaccine prepared from the live culture of fungi releasing MBS products has been shown.
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A 45 kDa protein was isolated from a soluble vaccine prepared from formaldehyde-killed spherules of Coccidioides immitis. From the N-terminal amino acid sequence, the protein yielded a 17-amino-acid peptide that was homologous to sequences of other fungal aspartyl proteinases. The coccidioidal cDNA encoding the proteinase was amplified using oligonucleotide primers designed from the 45 kDa N-terminal amino acid sequence and a fungal aspartyl proteinase consensus amino acid sequence. The PCR product was cloned and sequenced, and the remaining 5' upstream and 3' downstream cDNA was amplified, cloned, and sequenced. The cDNA encoding the coccidioidal aspartyl proteinase open reading frame was cloned and the fusion protein containing a C-terminal His-tag expressed in E. coli. The recombinant aspartyl proteinase was purified by immobilized metal affinity chromatography. This recombinant protein will be used for further studies to evaluate its antigenicity, including protective immunogenicity.
The studies reported in this review indicate that, whereas the expectations from the molecular approach help to excite and enlighten us, the realities suggest that even some less modern approaches may provide the necessary practical solutions to problems of serology and vaccination against coccidioidomycosis. Concurrent conduct of the two approaches should maximize the chances for success.
Two vaccines to treat phythiosis insidiosi in horses were evaluated in 71 Costa Rican horses between 1982 to 1988. One vaccine used a cell-mass (CMV) as antigen and the other a soluble concentrated antigen (SCAV). Both vaccines cured horses infected with Pythium insidiosum (p value approximately 14%). The age of lesions prior to vaccination was important in the response of the horses to immunotherapy. All horses with lesions 0.5 months or less in duration were cured regardless of the vaccine used. Horses with lesions two or more months old did not respond to either vaccine. The age of the horses did not have any influence on their response to the vaccinations. The CMV produced a prominent inflammatory reaction at the side of injection, while the SCAV gave a low inflammatory reaction. In addition, the CMV lost its effectiveness two to three weeks after its preparation. By contrast, the SCAV maintained its ability to cure horses even after 18 months. Immunotherapy using SCAV can thus be used as the vaccine of choice in early cases of equine cutaneous pythiosis insidiosi.
We have developed an oral vaccine using the blastococcoid form of Candida albicans with a murine model of oral candidiasis with infection-resistant (BALB/c) and infection-prone (DBA/2) mice. Previous studies had demonstrated that enhanced clearance was linked to increased production of both Th1 and Th2 cytokines (IFN-gamma and IL-4, respectively) in regional lymph nodes, and increased secretion of IFN-gamma and NO in saliva. Subsequent study using oral Lactobacillus acidophilus (LAVRI-A1) induced a local cytokine pattern and enhanced clearance of intra-oral C. albicans similar to that noted in animals given killed candida vaccine. Thus, the experiment was repeated with a 2 month gap between completion of a vaccine course and challenge with live C. albicans. Compared to control mice, those given the oral vaccine had a 4 log reduction in colonisation on day 2, associated with twice the levels of secretion of IFN-gamma and IL-4 from the regional node cells, five times the level of nitric oxide in saliva, and suppression of NO production with MMLA induced a 2 log increase in oral colonisation. Thus while both IFN-gamma and IL-4 appear to contribute to clearance of oral C. albicans, IL-4 appears to act through a paracrine enhancement of NO production. Studies in man are in progress to define similar surrogate markers of immune protection, prior to oral vaccine trial.
A laboratory-prepared killed Microsporum canis cell-wall vaccine was evaluated under conditions simulating an accidental infection of a cattery, by inoculating eight- to nine-week-old cats with the vaccine or with a placebo control. The vaccinated cats developed high titres of anti-dermatophyte IgG as measured by an ELISA, and a small cell-mediated response against M canis as measured by a lymphocyte blastogenesis assay, using a whole fungus extract. After being inoculated the cats were challenged by the introduction of an infected cat into the same room. All the vaccinated and control cats became culture-positive for M canis within four weeks of the introduction of the infected cat. Four of the six control cats and all the vaccinated cats developed lesions consistent with dermatophytosis within 16 weeks after exposure to the infected cat.
Epizootiologic studies were carried out after a special programme in four calf-fattening combines harbouring trichophytosis-affected animals. Tested was the effectiveness of a Soviet vaccina, Tphi 130 under this country's conditions. It was found that 20 per cent of the examined contact calves and 23.3 per cent of the survivals were carriers of T. faviforme. It was demonstrated that the vaccine conferred solid immunity against the Bulgarian strains of T. faviforme.
Experiments in BALB/c mice were carried out to study a number of immunomodulators in combination with vaccination using a recombinant yeast hepatitis B vaccine. The use of poludan, ridostine, amixine, larifan, myekonide significantly enhanced specific antibody production and at least doubled cell-mediated immune response by 14 and 28 days postvaccination. The above-mentioned immunomodulators are planned to be used in vaccination with the recombinant yeast vaccine against hepatitis B.
Heat shock proteins (hsp) were identified in many infectious agents as immunodominant antigens with a protective effect. Immunization of laboratory animals by selected representants of hsp60, hsp70 and hsp90 isolated from several pathogens induced protective host immunity and significantly reduced clinical manifestation of infection. The present study involves preparation of a recombinant protein vaccine and a DNA vaccine. Both vaccines were derived from the hsp60 of the dermatophyte, Trichophyton mentagrophytes. Challenge trials with evaluation of the protective effect of vaccination were performed on calves and guinea-pigs. Both vaccination procedures reduced, in statistically significant fashion, the clinical course of skin mycosis in calves experimentally inoculated with the dermatophyte, T. mentagrophytes. In experiments with guinea-pigs, increased protection was only seen with DNA vaccination. After DNA vaccine application, no paravaccination side-effects (granulomas at the injection site, changed total state of the animal) were observed. Only vaccination with the recombinant protein in calf's experiment induced specific serum antibodies. This observation indicates that antibodies are not associated with protection. In summary, DNA vaccine hsp60 is the most promising for prevention of bovine trichophytosis.
Peptide mimotopes of capsular polysaccharides have been proposed as antigens for vaccines against encapsulated pathogens. In this study, we determined the antibody response to and efficacy of P13, a peptide mimetic of the Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan (GXM), in mice that produce human antibodies. P13 was conjugated to tetanus toxoid (TT) or diphtheria toxoid (DT) and administered subcutaneously in Alhydrogel with or without CpG to mice transgenic for human immunoglobulin loci (XenoMouse mice) and expressing either immunoglobulin G2 (IgG2) (G2 mice) or IgG4 (G4 mice). Mice were vaccinated and revaccinated two or three times. The serum antibody responses of the mice to GXM and P13 and antibody idiotype expression were analyzed by an enzyme-linked immunosorbent assay. The results showed that both P13-TT and P13-DT were antigenic, inducing a mimetic response to P13 in both G2 and G4 mice, and immunogenic, inducing a mimotope response including VH3 (idiotype)-positive antibodies to GXM in G2 but not G4 mice. CpG led to higher titers of IgG to P13 and GXM in P13-TT-vaccinated G2 mice. C. neoformans challenge of P13-protein conjugate-vaccinated and control G2 mice induced anamnestic IgG- and VH3-positive responses to GXM and was associated with a significantly decreased risk of death and a prolongation of survival in P13-DT-vaccinated mice compared to phosphate-buffered saline-treated or protein carrier-vaccinated mice. These findings reveal that P13 elicited a human antibody response with VH3 expression in human immunoglobulin transgenic mice that has been observed for human antibodies to GXM and support the concept that peptide mimotope-based vaccines may hold promise for the treatment of C. neoformans infections.
A vaccine consisting of formaldehyde-killed spherules of Coccidioides immitis, previously shown to provide protection against development of lethal coccidioidomycosis in laboratory animals, was evaluated in humans. This double blind "Phase 3" study, conducted during the period 1980 to 1985, involved 2,867 healthy subjects with no history of coccidioidomycosis and negative skin tests. Randomized into vaccine (n = 1,436) or placebo (n = 1,431) groups, the former received three intramuscular injections of 1.75 mg (dry weight) of spherules, the latter received three injections of sterile 0.85% NaCl solution. Compatible clinical presentation with cultural or serologic findings permitted detection of coccidioidomycosis. Of those receiving vaccine, nine developed coccidioidomycosis and nine additional were suspected of having the disease. Of the group receiving placebo, 12 developed coccidioidomycosis, and 13 additional were suspected of having the disease. All cases and suspected cases were mild. Under the conditions of this study, a slight but statistically insignificant reduction in the incidence of coccidioidomycosis was noted in the vaccinated group compared with that in the placebo group, and there was no discernible difference in severity of disease between these groups.