Mold Remediation >> Mold In House

Since the cell wall of C. immitis is made up primarily of nonprotein macromolecules, it contains a large amount of material presumably nonantigenic for T lymphocytes. Therefore, the whole organism is not the ideal vaccine candidate. Ideally, one would like to vaccinate patients selectively only with antigens Mold In House that stimulate a protective T-cell-mediated immune response. 

These antigens have been difficult to identify, and a consensus on what they are does not exist. Various approaches have been used to obtain antigenic proteins. In one, Mold In House a lysate of arthroconidia (coccidioidin) or spherules (spherulin) was made (47). Alkali treatment has also been used to extract antigens from arthroconidia and spherules. 

Another approach has been to use C. immitis antigens obtained without extraction Mold In House or autolysis. The advantage of this method is that one should obtain reproducible preparations of intact proteins. Cole and co-workers (49) found that when the outer conidial wall was removed from arthroconidia, the organism released various proteins (called the soluble conidial wall fraction). 

This mixture of proteins was extraordinarily effective in stimulating the proliferation of C. immitis-immune T cells in mice. Another antigenic mixture is a membranous material consisting primarily of proteins Mold In House and lipids that the spherule phase of the organism spontaneously releases (the spherule outer wall). 

This spherule wall fraction has been shown to be an active antigen in T-cell-mediated immune responses in mice (50). All of these mixtures are heterogeneous and difficult to fractionate biochemically. This is probably due, at least in part, Mold In House to differences in glycosylation, which makes physically separating the proteins difficult. 

To resolve this problem, Galgiani and his colleagues deglycosylated the proteins from a toluene spherule lysate by using hydrogen fluoride (51,52). Although this treatment does remove all sugars, it is extraordinarily harsh and yields less than 10% of the initial protein, Mold In House with most of the protein forming an insoluble precipitate. 

Nevertheless, the resulting product reacts with reference antiserum to C. immitis in immunoelectrophoresis. This antigen also stimulated a proliferative T-cell response in patient lymphocytes but not in those of the control group (noninfected donors). Another way Mold In House to attack the problem of generating pure C. immitis antigens is to use molecular biologic techniques. 

The advantage to this approach is that once antigens are molecularly cloned, and the protein is expressed, an essentially unlimited source of completely defined antigen is available. Therefore, one would not have to repeatedly grow C. immitis, extract the antigen, Mold In House and purify it from a complex mixture. In addition, with the molecular approach, antigens could be delivered as part of a living vaccine system, should that be required to effectively immunize people against coccidioidomycosis. 

We believe that systematically identifying and evaluating C. immitis T-cell reactive antigens in experimental animals is a rational approach to the ultimate development of a vaccine. Our laboratories, Mold In House in collaboration with Garry Cole, have used a murine T-cell line that is specific for soluble conidial wall fraction antigens to identify one cloned fragment of a C. immitis protein (53). 

Recently, Mold In House genomic DNA clones coding for this protein have been identified and sequenced. Significant homology exists between this C. immitis antigen and the human enzyme 4 hydroxyphenylpyruvate dioxygenase (54). This protein has been expressed in bacteria and was found to elicit T-lymphocyte proliferative responses in mice immune to C. immitis. 

We are testing its efficacy as an experimental vaccine. With the exception of alkali extracted spherules (55) and whole killed spherules (45), none of the T-cell reactive antigens have been shown to be immunoprotective in experimental models. However, Mold In House it is reasonable to expect that some antigen, or mixture of antigens, will be found that can confer protective immunity in experimental animals. 

Molecular strategies are available to accomplish this task and are an important area of future research. Once a vaccine has been successfully tested in animals, another human vaccine trial would be feasible. Dr. Kirkland is associate professor of pathology and medicine; Mold In House Dr. Fierer is professor of medicine and pathology and head of the Division of Infectious Diseases, University of California, San Diego School of Medicine. 

Currently, they are focusing on the genetic determinants for resistance to infection and on identifying candidates for a coccidioidomycosis vaccine. Acknowledgment We are grateful to Director of the Kern County Public Health Laboratory, Mold In House and for sharing unpublished data with us. The experimental work done in our laboratories has been supported by National Institutes for Health grants and by the Research Service of the Department of Veterans Affairs.

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