The lipid A immunity system. I. Induction of heterophile antibodies by enterobacterial lipopolysaccharides and their lipid A component

Abstract

The lipid A immunity system Induction of heterophile antibodies by enterobacterial lipopolysaccharides and their lipid A componentMice were immunized either with lipopolysaccharides (LPS) or with the respective lipid A components. After four days an increase in the number of direct plaque-forming cells with complement-dependent antibody against sheep red cells was seen. Cross-reactions were observed with red cells of cattle, goats, horses and rabbits. All LPS investigated, of Shigella, Escherichia coli, and S and R form Salmonella, as well as the lipid A component evoked this immune response. We therefore conclude that all enterobacterial LPS induce an antibody which cross-reacts with SRC and that the lipid A component is responsible for this reaction. Evidence is presented that these effects, such as toxicity, fever, pyrogen tolerance, leucopenia/leucocytosis, nonspecific immunity, and tumor inhibition are mediated by lipid A-induced antibodies. Materials and methods BacteriaThe Salmonella cholerae suis strains were kindly supplied by H. Momenteau and L. LeMinor, International Salmonella Centre, Institut Pasteur, Paris, the Escherichia coli strains by F. Grskov, Statens Seruminstitut, Copenhagen, Shigella sonnei (phase 11) by M. Mulczyk, Institut Immunologii i Terapii, D&wiadczalnej, Breslau, Poland. R and S form Salmonella were provided by 0. Luderitz, Max-Planck-Institut fur Immunbiologie, Freiburg and by R. Rohde, Nationale Salmonella-Zentrale, Hygiene-Institut, Hamburg, respectively. The bacteria were kept at 4 O C on peptone agar. 2.2. Preparation of antigens Abbreviations: SRC: Sheep red blood cell(s) PFC: Direct antibody plaque-forming cell(s) LPS: Lipopolysaccharide(s) PBS: Phosphate buffered saline MBB: Modified barbital buffer i.p.: intraperitoneal 2.2.1. Lipopolysaccharides Bacterja were grown in 100 liter steel containers for 30 h at 37 C, slightly aerated, with either modified Fraser's medium [20], or in synthetic medium: 0.1 M Tris, pH 7.4; 0.0005 M KH2P04; 0.002 M sodium citrate, pH 6; 0.02 M NH4C1; 0.001 M MgS04; 1 x M FeCI3; 1 x M W.-R. Rank, R. DiPauli and U. Flie-Rank Eur.

The Use of Bacterial Lipopolysaccharides to Show That Two Signals Are Required for the Induction of Antibody Synthesis

“…We stress that in normal spleen cultures :~ Identical cultures contained either 3 )< 106 SRBC or 3 X 106 HRBC, All cultures were assayed on day 4 using the erythrocytes indicated in the plaque-forming assay and each figure represents the mean of duplicate cultures. (23)(24)(25) and in adult mice (26) (d) A background immune response to TNP is also found. When nude spleen cells are cultured with different concentrations of LPS and assayed at various times using HRB C or TNP-HRBC as the indicator in the plaque-forming assay, PFC specific for both HRBC and TNP can be distinguished.…”

“…(a) LPS stimulates, in the absence of added erythrocytes or TNP determinants, a low level response that is greatly enhanced by addition of the given antigen (Table I; [26]). This in itself should occasion no surprise since the culture medium containing bovine serum is likely to have in it cross-reacting substances that like monovalent haptens are rendered immunogenic by LPS.…”

Immune and mitogenic responses by BALB/c, C3H/HeJ, and nude mice to Brucella abortus bacterin and lipopolysaccharide

“…In 1960 Redfearn reported that BA-LPS could be recovered from the phenol phase of the Westphal extraction procedure; endotoxin extracted from the Enterobacteriaceae has long been known to partition into the aqueous phase. Thus, experiments based on extracted materials incorrectly thought to be the Brucella endotoxin indicated that BA-LPS was not cytotoxic, pyrogenic, or antigenic (5,12,21,37). We have, therefore, reexamined the immunogenic and mitogenic properties ofbiologically active BA-LPS.…”

“…The antibody responses of normal BALB/c and C3H/HeJ mice to BA and BA-LPS were qualitatively and quantitatively similar. Both BA and BA-LPS were mitogenic for spleen cells from normal and nude BALB/c and C3H/HeJ mice but not for thymus cells from normal BALB/c or C3H/HeJ mice, suggesting that both preparations are B-cell mitogens.There are conflicting reports (5,12,16,21,37) about the ability of the brucellae to elicit the biological and immunological responses evoked by the Enterobacteriaceae. These early reports, based on various animal models, either confirmed or denied the cytotoxic, pyrogenic, and antigenic properties of killed Brucella abortus cells (BA) or B. abortus lipopolysaccharide (BA-LPS).…”

“…There are conflicting reports (5,12,16,21,37) about the ability of the brucellae to elicit the biological and immunological responses evoked by the Enterobacteriaceae. These early reports, based on various animal models, either confirmed or denied the cytotoxic, pyrogenic, and antigenic properties of killed Brucella abortus cells (BA) or B. abortus lipopolysaccharide (BA-LPS).…”