Characteristics of Mycoplasma hominis adhesion

Abstract

The high concentrations of sulfated glycolipids in the human endometrium (6) and in mammalian male germ cells (10) suggest an advantage for organisms that can use these glycolipids as receptors. Lingwood and coworkers recently found that M. hominis binds to 2 ,ug of sulfogalactoglycerolipid and sulfogalactosylceramide, or sulfatide, on thin-layer plates (7). However, it was not demonstrated that this attachment is a specific adhesin-receptor interaction rather than some form of nonspecific, perhaps ionic, binding.To better understand the ability of M. hominis to colonize and cause disease, we screened a variety of glycoconjugates for the ability to serve as receptors for M. hominis adherence. We also examined the specificity and avidity of M. hominis attachment to sulfatide.All saccharides, polysaccharides, and glycoproteins were purchased from Sigma Chemical Company (St. Louis, Mo.), as were cholesterol, cholesterol sulfate, phosphatidylcholine, and bovine serum albumin (BSA). The glycolipids GM1, asialo-GM1, GD1a, GDib, GT1b, GM2, GM3, galactosylceramide, glucosylceramide, lactosylceramide, globoside, and sulfatide were also obtained from Sigma. Asialo-GM2, trihexosylceramide, and Forssman antigen were purchased from BioCarb Inc., Gaithersburg, Md.M. hominis 1620 (passage level 3 to 5) was used predominantly in this study. Strain 1620 was originally isolated from the synovial tissue of a chronically infected septic arthritis patient (13). For comparison where noted, reference strain PG21, a rectal isolate which has been passed in the labora-* Corresponding author. tory for over 30 years, was used in some studies, as was patient blood isolate strain H5488 at passage level 3. All cultures were grown and metabolically labeled with [3H]palmitic acid (12 to 17 Ci/mmol; Dupont-New England Nuclear) as previously described (2). After harvesting and two washes in 0.05 M Tris-HCl (pH 7.6)-110 mM sodium chloride (TBS), the organisms were suspended in 10 ml of buffer, passed four times through a 23-gauge needle, and further diluted with buffer to approximately 108/ml. Depending on the experiment, this dilution provided approximately 106 cpm/ml.Thin-layer chromatography (TLC) overlays were performed as a modification of the method described in reference 5. Briefly, glycolipids and lipids were separated on aluminum-backed silica gel high-performance plates (Merck, Darmstadt, Germany) with chloroform-methanol-water (60: 35:8). Following chromatography, the plates were blocked with 1% BSA in TBS (TBS-BSA) for 1 h. Plates were then incubated for 3 h at 370C with approximately 108 CFU of unlabeled M. hominis per ml in TBS-BSA. After 30 min of washing in TBS, the plates were incubated overnight at 40C with monoclonal antibodies (1:1,000 dilution) specific for surface-exposed, protease-sensitive, integral membrane proteins of M. hominis (11). After another wash in TBS-BSA, the plates were incubated for 1 h at room temperature with

Members of the 70 kDa heat shock protein family specifically recognize sulfoglycolipids: Role in gamete recognition and mycoplasma‐related infertility

“…Other studies have reported the same receptor binding specificity for M. pneumoniae (Krivan et al, 1988) and confirmed the glycolipid binding specificity of M . homznis (Olson and Gilbert, 1993). Sulfoglycolipids were recently demonstrated as receptors for M. hyopneumoniae (Zhang et al, 1994a,b) as we would predict from our present studies.…”

“…Four speciesspecific mycoplasmas associated with infertility bind to SGC and SGG (Lingwood et al, 1990a,b). Other investi- gators have demonstrated a similar glycolipid binding specificity for Mycoplasma pneumoniae (Krivan et al, 1988) and sulfoglycolipids were confirmed as the only receptor species for Mycoplasma hominis (Olson and Gilbert, 1993). Mycoplasma hyopneumoniae was recently shown to bind SGC and sulfoglycolipids found to mediate the attachment of this organism to ciliated respiratory epithelium (Zhang et al, 1994a).…”

Current Status of the Mollicute (Mycoplasma) Lung Disease: Pathogenesis, Diagnostics, Treatment and Prevention

“…It is worthy to note that the apical microvillar border and cilia of the epithelium express the sialoglycoconjugate-type receptors, thus allowing the selective attachment of M. pneumoniae to the ciliated cells [Krivan et al, 1988Roberts, D.D. et al, 1989;Olson & Gilbert, 1993;Rottem & Naot, 1998]. Cytadhered M. pneumoniae is able to cause damage through generation of reactive oxygen species (ROS), which act in concert with host's endogenous ROS to induce oxidative stress.…”

Binding of mycoplasmas to solid phase adsorbents

“…It is likely that surface proteins and lipids that facilitate the adhesion process will have an effect on invasion (Krivan et al, 1989;Olson and Gilbert, 1993;Zhang et al, 1994;Athamna et al, 1996). It has been shown that bacterial and viral invasion is based on the ability of several pathogens to attach to fibronectin or sulphated polysaccharides (Christensen et al, 2001).…”