The activities to induce TNF-α production by a monocytic cell line, THP-1, and ICAM-1 expression and IL-6 production by human gingival fibroblasts were detected in plural membrane lipoproteins of Mycoplasma salivarium. Although SDS-PAGE of the lipoproteins digested by proteinase K did not reveal any protein bands with molecular masses higher than approximately10 kDa, these activities were detected in the front of the gel. A lipoprotein with a molecular mass of 44 kDa (Lp44) was purified. Proteinase K did not affect the ICAM-1 expression-inducing activity of Lp44, but lipoprotein lipase abrogated the activity. These results suggested that the proteinase K-resistant and low molecular mass entity, possibly the N-terminal lipid moiety, played a key role in the expression of the activity. The N-terminal lipid moiety of Lp44 was purified from Lp44 digested with proteinase K by HPLC. Judging from the structure of microbial lipopeptides as well as the amino acid sequence and infrared spectrum of Lp44, the structure of the N-terminal lipid moiety of Lp44 was speculated to be S-(2, 3-bisacyloxypropyl)-cysteine-GDPKHPKSFTEWV-. Its analogue, S-(2, 3-bispalmitoyloxypropyl)-cysteine-GDPKHPKSF, was synthesized. The lipopeptide was similar to the N-terminal lipid moiety of Lp44 in the infrared spectrum and the ICAM-1 expression-inducing activity. Thus, this study suggested that the active entity of Lp44 was its N-terminal lipopeptide moiety, the structure of which was very similar to S-(2, 3-bispalmitoyloxypropyl)-cysteine-GDPKHPKSF.

Saliva samples collected from 393 subjects with and without oral diseases were examined for concentrations of mycoplasmas and Mycoplasma species. Mycoplasmas were isolated from 383 (97%) of the 393 subjects. Viable counts ranged from zero to 7.6 x 107 CFU/ml (median, 6.9 x 104) and were significantly (P < 0.01) higher in diseased subjects, except for those with arthrosis temporomandibularis, than in controls. Of 1,400 isolates, 897 (64%), 442 (30%), and 8 (1%) were identified as Mycoplasma salivarium, M. orale, and M. hominis, respectively, and the remaining 73 isolates (5%) were unidentifiable. More than two-thirds of the isolates from diseased subjects versus only half from controls were identified as M. salivarium. In diseased subjects other than those with ostitis (especially those with arthrosis temporomandibularis), the incidence of M. salivarium was higher than that of M. orale, whereas the former occurred about as frequently as the latter in the controls. Antibodies to M. salivarium were also measured in sera from some subjects by the metabolism inhibition test. Sera with metabolism inhibition titers of 16 or greater were rated positive. There was no significant difference in the prevalence of antibodies between diseased subjects (60%) and controls (40%), but the mean titers (97 to 220) of all positive sera from diseased subjects were two to four times those for sera from controls. In addition, a fourfold or greater rise or fall of antibody titers to the organism was shown in paired sera from some subjects. On the basis of these results, M. salivarium was strongly suggested to participate etiologically in some cases of oral infection.

No abstract

Detection ofgranulocyte colony-stimulating factor (G-CSF), one of the substances responsible for proliferation and diEferentiation of granulocytes, has been performed up to the present by use of the granulocyte colony-formation assay, because of the lack ofa specific anti-G-CSF antibody. This has prevented the advancement of biological investigations of cell dynamics linked to G-CSF, e.g., cell localization of G-CSF and its pathophysiobogical changes. In the present work, two monoclonal antibodies (MAb), 1E7 and 4A6, against recombinant human G-CSF (rhG-CSF) were devel-o_ by cell hybridization between NS-1 myeboma cells and splenocytes from a mouse immunized with rhG-CSF. 1E7 and 4A6 were shown to be reactive with hG-CSF but not with other CSF (hGM-CSF, hIL-3, and mouse GM-CSF) by Western blot analysis.

S U M M A R YMycoplasmas which metabolized urea and produced small colonies on agar medium were isolated from the genital tracts of dogs and from the throats of squirrel monkeys. Attempts to isolate similar organisms from baboons, cats, horses, pigs and rabbits were not successful. The biological and physical properties of the mycoplasmas isolated from dogs and monkeys were closely similar to those of T-mycoplasmas isolated previously from man and cattle, so that the canine and simian strains could be regarded undoubtedly as members of the T-mycoplasma group. The metabolic-inhibition technique was used to show that the T-mycoplasmas of the four 'host' species were not related to known large colony-forming mycoplasmas of these species. The same technique revealed that some of the Tmycoplasma strains isolated from a particular species were different from one another, except the simian strains which appeared to be the same as each other. In addition, the results of tests with a single or a few strains isolated from each of the four species showed that the strains of one species were not serologically related to those of another. On the other hand, polyacrylamide gel electrophoresis studies, in which at least 1 1 lines were observed with T-mycoplasma antigens, indicated that the structural proteins of strains isolated from the various species were closely similar. The fact that all the strains examined could be regarded as belonging to one closely related group is discussed in the context of the eventual nomenclature for these organisms.