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Ureaplasma urealyticum is a common commensal of the female lower urogenital tract, yet it has been shown to be an important cause of chorioamnion infection, respiratory and central nervous system disease, and death in premature infants. It has been suggested that only certain serovars are capable of producing invasive disease. However, we previously showed that many serotypes are invasive and that perhaps antigen variability and host factors are more important determinants of ureaplasma infections than are different serotypes per se. The molecular characterization in this report describes a mechanism available to ureaplasmas for producing antigen variation. That antigen, designated MB and previously identified on U. urealyticum, contains serovar-specific and cross-reactive epitopes, is produced both in vitro and in vivo, is a predominant antigen recognized during ureaplasma infections of humans, undergoes a high rate of size variation in vitro, and is size variable on invasive ureaplasma isolates. In the present study, we cloned and sequenced the gene of the MB antigen from serovar 3, the serovar most commonly isolated from humans. The 3' two-thirds of the gene was shown to contain identical 18-nucleotide tandem repeats. PCR analysis and direct sequencing of two variants indicated that alterations within this repeat region are responsible for the size variation of the MB antigen. Intact recombinant serovar 3 MB antigen and truncated products, expressed by coupled in vitro transcription and translation of the cloned gene, were immunoprecipitated by both a serovar-specific monoclonal antibody and the serum of a U. urealyticum-infected patient, and these results identified the repeat region of the MB antigen as serovar defining. Resolution of the precise amino acids responsible for specific epitopes and characterization of similar genes in the other serovars should yield reagents useful in elucidating the role of antigen size variants in disease production and the role of specific antibody in protection from ureaplasma disease.

Monoclonal antibodies against Ureaplasma urealyticum serotype 4 were produced by immunizing BALB/c mice with whole-cell antigens of the U. urealyticum serotype 4 reference strain. Ten monoclonal antibodies differentiated into two groups were found: one group included five monoclonal antibodies recognizing a band in immunoblotting that had a molecular mass of 81 kDa, and a second group included another five monoclonal antibodies recognizing three bands in immunoblotting that had molecular masses of 81, 75, and 71 kDa. Fifteen clinical U. urealyticum isolates were selected for serotyping with serotype 4-specific monoclonal antibodies and polyclonal antisera 1 to 14. The results obtained with polyclonal and monoclonal antibodies suggest the existence of heterogeneity of the serotype antigens among clinical isolates of U. urealyticum serotype 4.

Ureaplasma urealyticum has previously been isolated from the cultured cerebrospinal fluid of 13 of 418 newborn infants; additional bloodstream isolates were obtained from the same population. Ten of the 13 cerebrospinal fluid and 3 bloodstream isolates were available for serotyping in the present study. By the use of serotype-specific reagents, including monoclonal antibodies, 70% of the cerebrospinal fluid isolates were identifiable as serotype 1, 3, 6, 8, or 10; i.e., they represented 5 of the 14 established serotypes or both presently defined genomic clusters. One of the bloodstream isolates was identified as serotype 3. Our data support the hypothesis that the property of invasiveness for unreaplasmas is likely not limited to one or a few particular serotypes among the 14 established serovars. Additionally, our study has shown that even in isolates of the same serotype, there can be size variation in the antigens expressed. Therefore, it would appear that many serotypes are invasive and that perhaps antigen variability and host factors may be more important determinants for ureaplasma infections than different serotypes per se.