Duodeno-gastric reflux and its consequences after resection of stomach by Billroth II and Roux methods

Monoclonal antibodies to surface determinants of V. cholerae R forms (R-McA) were obtained. R-McA and monoclonal antibodies to lipopolysaccharide (LPS) of V. cholerae S forms (S-McA) were used to show that the LPS of deeply altered vibrios, agglutinating only with RO serum, completely lost its O-side chain. Some common O determinants on the basis of S-McA were detected in typical cultures of V. cholerae O1 and RO vibrios which agglutinated to 1/4 T with O serum and, in low titers, with RO serum. V. cholerae O1 were not capable of specifically binding with R-McA. Not all R strains under study were identified with the use of available R-McA due to essential differences of their terminal monosaccharides responsible for serological specificity.     

Immune mechanisms and protective antigens of Vibrio cholerae serogroup O139 as a basis for vaccine development

We have characterized 11 isolates of Vibrio cholerae O139 Bengal with regard to properties deemed to be relevant for development of a vaccine against O139 cholera. For most strains two colony variants, A and B, which are nonhemolytic and hemolytic, respectively, were detected on blood agar. The A and B variants were associated with high- and low-level production of soluble hemagglutinin-protease, respectively. However, on Luria-Bertani agar both types formed opaque colonies, which has been shown to be associated with capsule formation. Interestingly, under the stationary tube-shaken flask culture conditions in yeast extract-peptone water medium which were used to stimulate the production of cholera toxin (CT) and toxin-coregulated pili, B variants constitutively produced CT and TcpA, two ToxR-regulated proteins, at 28 and 37 degrees C, whereas the production of these proteins by A variants was downregulated at the higher temperature. One of the strains, 4260B, having a well-exposed O antigen and capsule and the capacity to produce large amounts of TcpA, CT, and mannose-sensitive hemagglutinin pili but minimal amounts of the proteolytic soluble hemagglutinin, was selected to produce antibacterial antisera and as a challenge strain in protection studies using the rabbit ileal loop model. Rabbit antisera to live, heat-killed, or formalin-killed O139 vibrios or to purified O139 lipopoly-saccharide (LPS) as well as monoclonal antibodies (MAbs) to O139 LPS agglutinated all O139 isolates. However, when A and B variants of strain 4260 were tested for sensitivity to vibriocidal activity of these antibody preparations, only the B variant was killed. All of the antisera against live or killed O139 vibrios conferred passive protection against fluid accumulation induced by the challenge strain. The protective effects of the antisera were correlated to anti-LPS antibody titers rather than to titers against whole bacteria that had been grown for toxin-coregulated pilus expression. This protection was considerably higher than that conferred by antisera to classical, EI Tor, or recombinantly produced (classical) CT or CTB. Furthermore, MAbs to O139 LPS and CTB-CT exhibited a strong synergistic protection against O139 challenge irrespective of the level of sensitivity of challenge strains to O139 LPS MAbs in vibriocidal assays in vitro.     

Decoration of lipopolysaccharide with phosphorylcholine: a phase-variable characteristic of Haemophilus influenzae

Choline, although not a nutritional requirement for Haemophilus influenzae, is taken up from the growth medium and incorporated into its lipopolysaccharide (LPS). Incorporated choline is in the form of phosphorylcholine (ChoP) based on the reactivity with the monoclonal antibody with specificity for this structure, TEPC-15. Incorporation of [3H]choline from the growth medium and expression of the TEPC-15 epitope undergo high-frequency phase variation, characteristic of other LPS structures in this species. The expression and phase variation of ChoP require a previously identified locus involved in LPS biosynthesis, lic1. The first gene in lic1, licA, contains a translational switch based on variation in the number of intragenic tandem repeats of the sequence 5'-CAAT-3'. The full-length LicA polypeptide resembles choline kinases of eucaryotes, suggesting that the pathway for choline incorporation into the H. influenzae glycolipid has similarities to the pathway for choline incorporation in eucaryotic lipid synthesis. The display of ChoP, a host-like structure, renders the organism more rather than less susceptible to the bactericidal activity of human serum. The increased serum sensitivity of variants with ChoP correlates with higher serum immunoglobulin G titers to LPS containing this structure. ChoP appears to be a cell surface feature common to a number of pathogens of the human respiratory tract, including Streptococcus pneumoniae and mycoplasmas. In the case of H. influenzae, its primary contribution to pathogenesis does not appear to be antigenic variation to evade host humoral clearance mechanisms.     

Human antibody response to Helicobacter pylori lipopolysaccharide: presence of an immunodominant epitope in the polysaccharide chain of lipopolysaccharide

We have examined the antibody response to Helicobacter pylori lipopolysaccharides (LPS) in humans. We used sera from patients with gastroduodenal diseases and healthy adults infected or not infected with H. pylori. Data from the experiments for antibody binding to LPS suggested that the polysaccharide chains from many H. pylori strains showed high immunogenicity in humans. Sera from most (above 70%) H. pylori-infected individuals contained immunoglobulin G (IgG) antibodies against the polysaccharide region highly immunogenic H. pylori LPS. The IgG titers of individual serum samples that reacted strongly with highly immunogenic LPS were quite similar (r2 = 0.84 to 0.98). The results suggest wide distribution among H. pylori strains of a highly antigenic epitope in the polysaccharide moieties of their LPS. Also, the similarity in the titers of individual serum samples against highly immunogenic LPS points to the existence of epitopes sharing a common structural motif. However, some strains showed low antigenicity, even those with polysaccharide-carrying LPS. The dominant subclass of IgG that reacted with the highly immunogenic LPS was IgG2, which was preferentially raised against polysaccharide antigens. Recently, a structure that mimics that of the Lewis antigens was identified in the O-polysaccharide fraction of H. pylori LPS; however, no correlation between antigenicity of the polysaccharide chain in humans and the presence of Lewis antigens was found. The IgA and IgM titers against H. pylori LPS seemed to be mostly nonspecific and directed against lipid A. In a few cases, however, sera from individuals infected with H. pylori gave strong IgA and IgM titers against the highly immunogenic polysaccharide. In conclusion, the LPS of many H. pylori strains possess an antigenic epitope in their polysaccharide regions that is immunogenic in humans. However, our results show that the antigenic epitope is unlikely to be immunologically related to structures mimicking Lewis antigens.     

The lipopolysaccharide of Porphyromonas gingivalis is not antigenically cross-reactive with that of other species

Large numbers of Porphyromonas, Prevotella, and Bacteroides strains were screened by 3 monoclonal antibodies (MAbs) and 8 rabbit antisera raised against Porphyromonas gingivalis, in order to detect any possible recognition of non-P. gingivalis surface antigens by these immunoreagents. All three MAbs, which were LPS-specific, extensively recognized LPS from 10 P. gingivalis strains in immunoblotting, whereas they recognized none of the 34 non-P. gingivalis strains. Rabbit antisera were similarly specific for P. gingivalis cells in immunofluorescence and with LPS in grid-blotting, but several of them recognized LPS from one Prevotella melaninogenica and 5 Prevotella intermedia strains in Western blotting. Since several pre-immune sera and an irrelevant serum raised to a Streptococcus species recognized up to 5 of these preparations, we exclude that the reactions were due to antigens shared by P. gingivalis and Prevotella. Rather, we consider that they were false-positive reactions due to natural antibodies, stimulated in a non-specific manner upon immunization with P. gingivalis, in animals whose immune systems were sensitized to Prevotella species before immunization.     

alpha-GlcNAc-1-->2-alpha-glc, the Salmonella homologue of a conserved lipopolysaccharide motif in the Enterobacteriaceae, elicits broadly cross-reactive antibodies

To define cross-reactive epitopes in Salmonella lipopolysaccharide (LPS), antisera designated anti-S, anti-Ra, and anti-Re were generated against smooth (S), complete-core (Ra), and deep-core mutant (Re) strains, respectively, and characterized immunochemically. The reactivities of anti-Ra and anti-S with rough LPS (rLPS) chemotypes in enzyme-linked immunosorbent assays (ELISA) decreased progressively with increasing truncation of the complete-core oligosaccharide (e.g., Ra > Rb1 >.Re), while that of anti-Re increased (Ra < Rb1 <.Re). Anti-Ra was relatively more reactive with nonhomologous smooth LPS (sLPS) than anti-S, which in turn was more reactive than anti-Re. This order reflected the relative reactivities of these sera with outer-core rLPS but not those with inner-core rLPS, which suggests that the cross-reactivities of all three sera with sLPS were mediated by antibodies which bind outer-core determinants. Anti-Ra, but not anti-S or anti-Re, reacted with molecules substituted by O chains in immunoblots and revealed ladder-like patterns in sLPSs of various serospecificities. Anti-Ra, however, did not react with O-antigen-specific neoglycoconjugates in ELISA, thus demonstrating specificity for core epitopes. Ra and Rb1 but not other Salmonella core chemotypes inhibited the reactivity of anti-Ra with sLPS in ELISA, which showed that the terminal outer-core disaccharide, alpha-GlcNAc-1-->2-alpha-Glc (GlcNAc-->Glc), was the major epitope of cross-reactive antibodies in the serum. GlcNAc-->Glc represents the conserved motif alpha-hexose-1-->2-alpha-hexose in cores of the Enterobacteriaceae, other homologues of which should likewise be cross-reactive. These results demonstrate that S or Re strains do not elicit cross-reactive antibodies and indicate that immunization with Ra strains may represent a general strategy for eliciting cross-reactive antibodies against LPSs from enteric bacteria.     

Production and characterization of a set of mouse-human chimeric immunoglobulin G (IgG) subclass and IgA monoclonal antibodies with identical variable regions specific for Pseudomonas aeruginosa serogroup O6 lipopolysaccharide

The heavy- and light-chain variable regions from a murine monoclonal antibody that recognize Pseudomonas aeruginosa serogroup O6 lipopolysaccharide (LPS) were used to generate a series of chimeric mouse-human monoclonal antibodies with identical variable regions. The murine variable-region gene segments were cloned into an immunoglobulin (Ig) cDNA expression vector that contained the human kappa light-chain and IgG1 constant regions. The IgG1 heavy-chain constant region was then replaced with the human IgG2, IgG3, IgG4, or IgA1 heavy-chain constant region. The five different expression vectors were transfected into Chinese hamster ovary cells for antibody production. The chimeric antibodies exhibited immunoreactivity and affinity similar to that of the parental murine IgG antibody toward whole cells of a serogroup O6 strain. In vitro complement deposition assays demonstrated that the chimeric IgG4 and IgA antibodies did not mediate the deposition of complement component C3 onto the surface of either purified LPS or whole bacteria. The chimeric IgG1 and IgG3 antibodies were similar in their ability to deposit C3 onto the surface of both bacteria and LPS, while IgG2 antibody was more effective at depositing C3 onto the surface of bacteria than onto purified LPS. The pattern of opsonophagocytic activity of the chimeric monoclonal antibodies was similar to that of complement deposition onto bacterial cells in that the chimeric IgG1 and IgG3 had the highest opsonic activity. Although IgG2 deposited more C3 onto the bacterial surface than did IgG4 or IgA, all three of these isotypes had low opsonic activity against the serogroup O6 target strain. This series of related antibodies will help reveal functional differences in efficacy among protective antibodies to P. aeruginosa and will be critical for defining the optimal formulation of either a vaccine for active immunization or a polyclonal intravenous IgG or monoclonal antibody cocktail for passive immunotherapy.     

Opsonization of Actinobacillus actinomycetemcomitans by immunoglobulin G antibodies to the O polysaccharide of lipopolysaccharide

Sera of localized juvenile periodontitis (LJP) patients colonized by Actinobacillus actinomycetemcomitans serotype b often contain markedly elevated levels of immunoglobulin G (IgG) antibodies to serospecific determinants in the O polysaccharide of lipopolysaccharide (LPS), as well as to outer membrane proteins of this species. IgG antibodies in LJP sera are known to opsonize A. actinomycetemcomitans for subsequent phagocytosis and killing by human neutrophils. The objective of this study was to determine whether outer membrane proteins or serospecific determinants in LPS are the primary target for opsonic IgG antibodies in LJP sera. An A. actinomycetemcomitans serotype b O-polysaccharide affinity column was constructed and subsequently used to purify LPS-specific IgG antibodies from LJP serum. The affinity-purified anti-LPS IgG antibodies were enriched in content of IgG2 (66.2%, compared with 37.0% in the total IgG fraction) and were immunospecific for A. actinomycetemcomitans serotype b LPS. In an opsonophagocytic assay using neutrophils from donors who were homozygous for the H131 allotype of Fcy receptor IIa (CD32), it was found that LPS-specific IgG antibodies exhibited substantially greater opsonic activity toward A. actinomycetemcomitans serotype b than an LJP IgG fraction that was depleted of LPS-reactive antibodies but contained antibodies against outer membrane proteins of this species. The results of this study indicate that serospecific determinants in the O polysaccharide of A. actinomycetemcomitans serotype b are a principal target for opsonic antibodies in sera of LJP subjects.     

An analysis of the central pathway of vestibulo-sympathetic responses

The sera of 159 patients with monoclonal gammopathies were examined for the presence of anti-thyroglobulin (Tg) activity. An enzyme-linked immunosorbent assay was employed. Thirty-one (19.5%) sera were found to bind Tg. The activity against Tg was further confirmed by using purified immunoglobulins and employing competition assays. The anti-Tg antibodies were found in the sera of patients with IgG, IgM and IgA gammopathies. Anti-Tg antibodies were more frequent among patients with IgG gammopathy. Autoantibodies to Tg are found in patients with Hashimoto's thyroiditis, Graves' disease and occasionally in patients with thyroid carcinoma. Natural autoantibodies directed against human Tg have been detected, as well, in healthy subjects. None of the patients in the present study whose serum was found to contain high titers of anti-Tg human monoclonal antibodies had any clinical or biochemical evidence of thyroid disease. Our results of a high incidence of anti-Tg activity in the sera of patients with monoclonal gammopathies support previous reports of autoantibody properties characteristic of these immunoglobulins.     

Complement, opsonins, and the immune response to bacterial infection in burned patients

Studies were performed to evaluate complement, opsonins, and the immune response to bacterial infection in burned patients. Concentrations and functional acitivities of components of the classical and alternative complement pathways were measured in the sera of four septic, two bacteremic, and four nonseptic burned patients. In addition, heat-labile and heat-stable opsonic activity and agglutinin titers directed against the infecting bacterial strains were measured in the sera of the four septic patients and in an additional group of 11 septic burned patients with abnormal complement profiles. Functional activity of the alternative complement pathway and the concentration of properdin were shown to be persistently decreased during eight weeks postburn in the septic, bacteremic, and nonseptic burned patients; reduced classical pathway activity was demonstrated during the initial postburn period only in the septic patients. Two of the 15 septic patients had decreased heat-labile serum opsonic activity for their infecting bacterial strains, which occurred only during the initial postburn period. Heat-stable opsonins and agglutinin titers in the patients' sera directed against the infecting bacterial strains were equivalent to those in normal human sera, except for the agglutinin titers to Streptococcus faecalis which were increased in the patients' sera in comparison to the normal sera. These results indicate that the multiple complement abnormalities which occur in septic burned patients do not predispose these patients to bacterial infection by decreasing serum opsonic activity. Moreover, heat-stable immune IgG antibodies are not produced during septicemia which facilitate opsonization of the infecting bacterial strains in the absence of an intact complement system.     

Immunological properties of Weil-Felix test negative sera from patients with Japanese spotted fever

Sera from 4 out of 19 patients with the Japanese spotted fever were negative to OX2 antigen of Weil-Felix (WF) test. These WF test negative sera were analyzed by ELISA and immunoblot used whole cells and lipopolysaccharides (LPS) of rickettsiae and Proteus strains as antigens. These acute-phase sera have already possessed the IgG antibodies against LPS of Proteus OX2 strain, whereas IgM antibodies in these acute- and convalescent-phase sera did not react with this LPS. On the other hand, the reactivity of IgM antibodies of the convalescent-phase sera in the 2 patients with LPS of Proteus OX19 strain increased as compared with that of the acute-phase sera by ELISA, and these IgM antibodies also showed the reactivity with bands of OX19-LPS in the immunoblot. On the basis of these results, it is interpreted that the WF test negative sera from patients with Japanese spotted fever are due to the presence of IgG antibodies against OX2-LPS in the sera.     

Randomized controlled trial of ipratropium bromide and frequent low doses of salbutamol in the management of mild and moderate acute pediatric asthma

The standardized enzyme-linked immunosorbent assay (ELISA) for measurement of serum immunoglobulin G (IgG) antibody responses to meningococcal C polysaccharide has been modified to employ assay conditions that ensure specificity and favor detection primarily of high-avidity antibodies. The modified and standard assays were used to measure IgG antibody concentrations in sera of toddlers vaccinated with meningococcal polysaccharide vaccine or a meningococcal C conjugate vaccine. The results were compared to the respective complement-mediated bactericidal antibody titers. In sera obtained after one or two doses of vaccine, the correlation coefficients, r, for the results of the standard assay and bactericidal antibody titers were 0.45 and 0.29, compared to 0.85 and 0.87, respectively, for the modified assay. With the standard assay, there were no significant differences between the geometric mean antibody responses of the two vaccine groups. In contrast, with the modified assay, 5- to 20-fold higher postvaccination antibody concentrations were measured in the conjugate than in the polysaccharide group. Importantly, the results of the modified assay, but not the standard ELISA, paralleled the respective geometric mean bactericidal antibody titers. Thus, by employing conditions that favor detection of higher-avidity IgG antibody, the modified ELISA provides results that correlate closely with measurements of antibody functional activity that are thought to be important in protection against meningococcal disease.     

Production of mouse monoclonal anti-idiotypic antibodies specific to epitopes of tumor-associated mucin TAG-12

The tumor-associated mucin-glycoprotein TAG-12 is strongly expressed in approximately 96% of all breast cancer patients and nearly 68% of all ovarian cancers. The experimental results of this work indicated that humoral immune response against TAG-12 is possible. Immunization with anti-idiotypic monoclonal antibodies produces this response. In this experiment, anti-idiotypic monoclonal antibodies represent the internal image of a specific epitope on TAG-12. Monoclonal antibody (MAb) 12H12 was selected to produce anti-idiotypic antibodies (anti-Ids) because of its high reactivity with TAG-12. Syngeneic murine anti-Ids were developed by immunization of BALB/c mice with the 12H12-Fab-KLH conjugate. A competitive assay with purified TAG-12 was utilized to identify anti-Ids with mirror image function. Two MAbs with "internal image" specificity were selected, 5H8 and 5H2. Two New Zealand White rabbits were immunized with 5H8. Serum samples tested 6 weeks after the initial immunization showed comparable titers against TAG-12. The binding capacities of the rabbit sera to different human breast as well as nonbreast cancer cell lines demonstrated strong binding with TAG-12-positive breast cancer cell lines. Competitive inhibition assays demonstrate that Ab3 and purified TAG-12 totally inhibit the binding of 12H12 antibody to TAG-12-positive cells. No inhibition was detectable with unrelated MAbs or normal mouse immunoglobulin. Binding assays with polyclonal Ab3 serum and several human cancer cell lines showed reactivity to nearly every tested cell line. Soluble TAG-12 showed no inhibition, indicating that this binding is due to a different set of idiotypes. Anti-Id 5H8 elicited an immune response to TAG-12. Utilization of anti-Id as a vaccine against the breast cancer-associated tumor antigen TAG-12 was successfully demonstrated in a xenogeneic animal model.     

Bovine mastitis caused by Listeria monocytogenes: kinetics of antibody responses in serum and milk after experimental infection

The kinetics of antibodies in serum and milk directed against proteins from Listeria monocytogenes were studied using 4 lactating cows after infection was experimentally induced in the udder with four strains of serotypes 4b or 1/2a. Antibodies (IgG and IgA) in samples of composite quarter milk and serum of the cow were measured by indirect ELISA. Microtiter plates were coated with proteins obtained from the culture supernatant of L. monocytogenes 4b. After challenge, an IgG response in serum and milk to listerial infections in the udder occurred for all cows, although the response varied among cows. In sera, the IgG titers reached a peak at 9 to 13 wk after challenge and remained elevated until 21 to 33 wk after challenge. In milk, the IgG titer increased significantly 3 wk after the challenge for all cows. A weak and nonpersistent increase in IgA antibodies also occurred. These results indicate that IMI by L. monocytogenes induced an increase of antibodies in milk, which could be detected with an ELISA test using our antigenic preparation. Therefore, this antigenic preparation could be used for the evaluation of a new method of diagnosis for bovine mastitis caused by L. monocytogenes.     

Characterization of lipopolysaccharide-deficient mutants of Pseudomonas aeruginosa derived from serotypes O3, O5, and O6

Well-characterized rough mutants are important for the understanding of structures, functions, and biosynthesis of lipopolysaccharide (LPS) in gram-negative organisms. In this study, three series of Pseudomonas aeruginosa LPS-deficient mutants, namely PAC strains derived from serotype O3, AK strains derived from strain PAO1 (serotype O5), and serotype O6-derived mutants were subjected to biochemical analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining as well as immunochemical characterization using LPS-specific monoclonal antibodies. The O-side-chain deficiency among the O6-derived mutants was also examined, and three mutants, A28, R5, and H4, were subsequently chosen for the elucidation of component sugars of the core structure of serotype O6 LPS. LPS of strain A28 has L-rhamnose and proportionally higher amounts of D-glucose, a feature shared by the O5-derived mutant, strain AK1401 (previously demonstrated as a mutant with a core-plus-one O repeat). In contrast strains R5 and H4 were shown to be devoid of L-rhamnose and have low and undetectable amounts of D-glucose, respectively, which indicated their core deficiency. The LPS-deficient or -sufficient characteristics of the P. aeruginosa strains examined correlated will with serum sensitivity data. This report represents a comprehensive analysis of rough mutants derived from O3 and O5 strains that have been used by others in many studies and a first look at the core oligosaccharide region of serotype O6 LPS obtained with the O6-derived mutants generated in this study.     

Application of transcranial Doppler sonography in surgical aspects of hypertensive putaminal haemorrhage

Serum antibodies from human immunodeficiency virus type 1 (HIV-1)-infected long-term non-progressors (LTNPs) and non-LTNPs were evaluated for virus neutralization and infection enhancement in vitro. Sera from LTNPs had higher average titers of neutralizing antibodies to HIV-1 strains IIIB and MN and more frequently neutralized primary isolates from progressors (14.9% vs. 1.3%, P = .002). Replication-competent HIV-1 was isolated from peripheral blood mononuclear cells and lymph nodes of 3 LTNPs. All viruses from LTNPs had a non-syncytium-inducing phenotype, were resistant to neutralization by autologous serum obtained at the time of virus isolation, and showed little evidence of a heightened sensitivity to neutralization by heterologous sera. Complement-mediated, antibody-dependent enhancement (C'-ADE) of HIV-1IIIB and primary isolates was equally prevalent for sera from LTNPs and non-LTNPs. Results indicate that LTNPs produce vigorous serum antibody responses and that long-term nonprogression is not associated with homologous neutralization or the absence of C'-ADE.     

Effects of azadirachtin on mortality, growth, and immunological function in the wolf spider, Schizocosa episina (Araneae: Lycosidae)

Broth culture supernatants from 14 (34%) out of the 41 H. pylori strains tested, induced vacuolization in Intestine 407 cells in titers ranging from 1:2 to 1:64. 20% of H. pylori strains isolated from children and 42% of strains isolated from adults expressed vacuolating activity. Serum antibody to cytotoxin produced by H. pylori was detected with a neutralization assay. Anticytotoxic antibodies were present in all sera from patients infected with cytotoxic H. pylori strains. The toxin-neutralizing activity of sera from individuals infected with H. pylori suggests that the cytotoxin is produced in vivo.     

Endothelial cell activation by sera containing HLA antibodies is mediated by interleukin-1

BACKGROUND: It has been suggested that antibodies which are associated with chronic pathological conditions such as chronic rejection and autoimmune diseases have the capacity to activate endothelial cells by induction and up-regulation of adhesion molecules. It has also been suggested that HLA antibodies formed by patients awaiting transplantation can activate endothelial cells. These antibodies include HLA and those that bind to endothelial cells. METHODS: We have further investigated this phenomenon using monoclonal antibodies against HLA class I determinants and sera from aortic valve graft recipients, containing strong HLA antibodies. The effect of 24-hr incubation of antibodies/serum with human umbilical vein endothelial cells (HUVECs) on adhesion molecule expression was measured by flow cytometry. RESULTS: HLA monoclonal antibodies had no effect on ICAM-1 expression on HUVECs. Five of 31 (16%) patients' sera caused strong up-regulation of adhesion molecules (ICAM-1, vascular cell adhesion molecule-1, and E-selectin) but this did not correlate with HLA specificity, IgG, or IgM binding to HUVECs. The activity, found in whole serum and IgG-depleted fractions was inhibited by neutralizing antibodies against interleukin (IL)-1beta and tumor necrosis factor-alpha. Examination of patient sera for presence of IL-1beta demonstrated high levels of IL-1beta in all five sera (range, 30 -500 U/ml) as well as in samples from an additional three patients. CONCLUSION: The ability to activate endothelial cells detected in our patient sera was caused by cytokines and not antibody. Our observation that addition of cytokines to sera before separation into large and low molecular weight fractions demonstrated retention of cytokines in both fractions may be a confounding issue when investigating endothelial cell activation by patients' sera.     

Detection of serum antibodies against Chlamydia pneumoniae by ELISA

Chlamydia pneumoniae causes pneumonia and other respiratory infections in children, adolescents and adults. We tried to evaluate the diagnostic value of detection of serum antibodies by ELISA for C. pneumoniae infections in Japanese children. Serum IgG, IgA and IgM antibodies to C. pneumoniae were determined by the microimmunofluorescence (MIF) test. Serum IgG and IgA antibodies were also determined by ELISA test kits. Results obtained by ELISA were compared with those obtained by MIF test. IgG antibody to C. pneumoniae was detected in 135 (39.5%) by ELISA and in 125 (36.5%) by MIF out of 342 sera from Japanese infants and children without respiratory infections (aged from 2 months old to 15 years old). IgA antibody to C. pneumoniae was detected in 129 (37.7%) by ELISA and in 117 (34.2%) by MIF out of 342 sera tested. Of 342 specimens 113 were IgG-positive by ELISA and MIF (sensitivity: 90.4%, specificity: 89.9%, r = 0.853). Of 342 sera 28 had IgG antibody titers of 1:256 and none had titers 1:512 or higher by MIF. Of 28 infants and children a total of nine were less than 4 years of age. On the other hand, of 342 specimens 99 were IgA-positive by ELISA and MIF (sensitivity: 84.6%, specificity: 86.7%, r = 0.769). Of 342 sera 16 had IgA antibody titers of 1:256 or higher by MIF. Of 16 infants and children, ten were less than 4 years of age. ELISA had excellent sensitivity and specificity relative to MIF test for detection of IgC and IgA antibodies to C. pneumoniae. It was suggested that C. pneumoniae infection in Japanese infants and children under 4 years of age was not infrequent.     

Studies on Neisseria gonorrhoeae strains from test-of-cure specimens. Correlation between the in vitro susceptibility to penicillin and the sensitivity to the complement-dependent bactericidal activity of normal and convalescent human serum

Sixty-seven out of 88 Neisseria gonorrhoeae strains isolated from test-of-cure (TOC) specimens during a five-months' period were included in the study. For 62 patients sufficient information was obtained in order to distinguish between relapse (34 ptt) and re-infection (28 ptt). For comparison with strains from these two groups of patients, 63 urogenital and 21 pharyngeal gonococcal strains isolated during the same period of time were randomly selected. The distributions according to susceptibility to penicillin for TOC strains and control strains corresponded to those found for the total number of TOC strains (275) and other strains (3,345) tested in 1979, respectively. The TOC strains did not differ from the control strains in sensitivity to the complement-dependent. The TOC strains did not differ from the control strains in sensitivity to the complement-dependent bactericidal activity of normal human serum. However, gonococcal strains less susceptible to penicillin in vitro (MIC values within the range 0.1-2.0 microgram/ml) were significantly more sensitive to the complement-dependent activity of normal human serum (P less than 0.01) than strains fully susceptible to penicillin (MIC less than 0.01 microgram/ml.) Penicillin-resistant strains (MIC greater than 2.0 microgram/ml) did not differ from strains susceptible to less than 0.1 microgram penicillin/ml and were slightly more serum-resistant than the less susceptible strains (P less than 0.05). No difference in serum-sensitivity of urogenital and pharyngeal isolates could be demonstrated. The level of bactericidal activity of homologous convalescent serum was unrelated to the presence of antibodies either to gonococcal pili or crude gonococcal antigen preparations. The sensitivity to normal human serum of a certain strain was not correlated with sensitivity to homologous convalescent serum.