Placental transfer of IgG antibodies specific to Klebsiella and Pseudomonas LPS and to group B Streptococcus in twin pregnancies.

Group B Streptococcus (GBS), Klebsiella spp. and Pseudomonas spp. are important aetiological agents of neonatal infections in Brazil. There is a lack of data in the literature regarding the specific transport of immunoglobulin G (IgG) against these pathogens in multiple pregnancies. Maternal (n = 55) and umbilical cord (n = 110) blood samples were prospectively collected at birth from 55 twin pregnancies. The factors associated with cord levels and transfer ratios of IgG against GBS, Klebsiella and Pseudomonas were examined. The IgG umbilical cord serum levels specific to GBS, Klebsiella LPS and Pseudomonas LPS were significantly associated with maternal-specific IgG concentrations and the presence of diabetes. The anti-Klebsiella IgG cord serum concentrations were also related to birthweight and the presence of hypertension. The transfer ratios against GBS and Pseudomonas LPS were associated with maternal-specific IgG concentrations. The transfer ratios for GBS and Pseudomonas LPS were associated with gestational age at delivery and the presence of diabetes, respectively. None of the examined parameters were related to Klebsiella LPS transfer ratios. We conclude that in twin pregnancies, specific maternal IgG serum concentrations and diabetes were the parameters associated with umbilical cord serum IgG concentrations reactive with the three pathogens investigated. All the other parameters investigated showed different associations with neonatal-specific IgG levels according to the antigen studied. There was no uniformity of the investigated parameters regarding association with placental IgG transfer ratios against the GBS, Pseudomonas LPS and Klebsiella LPS.

Antibody response to pneumococcal capsular polysaccharide vaccine in Down syndrome patients.

The majority of children with Down syndrome (DS) tend to have frequent bacterial infections including recurrent respiratory infections. Our objective was to evaluate the production of antibodies to pneumococcal polysaccharide antigens after active immunization in DS subjects. IgG antibodies to pneumococcal serotypes (1, 3, 6B, 9V, and 14) were measured before and 6 weeks after immunization with a 23-valent pneumococcal vaccine (Pneumo23, Pasteur-Merrieux) in 6- to 13-year-old DS children (N = 17) and in aged-matched normal controls (N = 30). An adequate response was defined as a 4-fold increase over baseline or a post-immunization level of specific pneumococcal serotype antibody > or = 1.3 microg/mL. After immunization, all DS children had an increase in post-immunization levels against all serotypes analyzed. A 4-fold or more increase was observed in all DS children concerning serotypes 1 and 14, in 90% of subjects for serotypes 3 and 9V, and in 65% for serotype 6B. Regarding this increase, 8 of the 17 DS children had an adequate response to all serotypes analyzed, 8/17 patients to 4 serotypes and 1/17 to 3 serotypes. However, when we compared post-immunization levels between DS children and controls, we observed lower levels in the former group (P < 0.05) for all serotypes except serotype 3. We conclude that pneumococcal polysaccharide immunization could be beneficial for these DS children.

Functional alterations of human neutrophils by medium-chain triglyceride emulsions: evaluation of phagocytosis, bacterial killing, and oxidative activity.

Medium-chain triglyceride (MCT) and long-chain triglyceride (LCT) emulsions currently used in nutritional therapy were evaluated for their in vitro effect on neutrophil oxidative metabolism, phagocytosis, and bacterial killing activities. Neutrophils from healthy adult male volunteers were assessed after blood incubation with commercially available fat emulsions containing LCT, MCT, or a mixture of 50% MCT and 50% LCT at a final triglyceride concentration of 20 mg/ml. It was observed that MCT-containing emulsions stimulated nitroblue tetrazolium (NBT) dye reduction by neutrophils as determined by a cytochemical NBT test performed directly on whole blood. This effect was dose dependent. However, after lipid removal by cell washing, the MCT-treated neutrophils showed decreased production of hydrogen peroxide (H2O2) and NBT reduction in response to bacterial lipopolysaccharide or phorbol myristate acetate stimuli as well as impaired phagocytosis and killing of Staphylococcus aureus. In contrast, the LCT emulsion did not alter any of the neutrophil functions evaluated. The present data suggest that MCTs elicit the oxidative metabolism of neutrophils, probably by phagocytosis of fat particles and, depending on the lipid concentration, this effect may not be reversible, leading to impairment of the cellular response to subsequent membrane stimuli.

Colostral neutrophils express Fc alpha receptors (CD89) lacking gamma chain association and mediate noninflammatory properties of secretory IgA.

Colostrum plays an important role in protecting newborn infants against acute gastrointestinal and respiratory infections. IgA antibodies have been considered the major effector component; however, the role of their receptors on colostral phagocytes, especially neutrophils, has not been studied. Here, we demonstrate that CD15+ colostrum neutrophils express IgA Fc receptors (Fc alphaR, CD89) at levels similar to those of blood neutrophils. Most colostral cells (70%) bear secretory IgA (SIgA) on their surface (and intracellularly), whereas blood cells do not. The Fc alphaR on colostral neutrophils was identified as the a.1 isoform with a similar molecular mass (55-75 kDa) as that identified for blood neutrophils. Removal of N-linked carbohydrates revealed a major protein core of 32 kDa for both cell types. In contrast, co-immunoprecipitation and immunoblot experiments using a mild detergent, digitonin, revealed a lack of gamma chain association with Fc alphaR (gamma-less) exclusively on colostral neutrophils. The functional role of these gamma-less Fc alphaR cells was evaluated by measuring superoxide release and killing of SIgA-coated enteropathogenic E. coli. No increase in superoxide release was observed in colostral cells compared with blood neutrophils, whereas optimal release was obtained with PMA stimulation. Furthermore, despite similar bacterial phagocytosis index between both cell types, IgA-mediated bacterial-killing was not detectable with colostral neutrophils, whereas killing was detectable on blood cells. These results reveal exclusive expression of gamma-less Fc alphaR on colostral neutrophils associated with receptor hyperoccupation by IgA and with low, bacterial-killing activity, which suggest that this receptor may mediate noninflammatory effects of SIgA.

Effect of therapy with recombinant human interferon-gamma on the release of nitric oxide by neutrophils and mononuclear cells from patients with chronic granulomatous disease.

The aim of this study was to investigate the effect of recombinant human interferon-gamma (rHuIFN-gamma) therapy on the release of nitric oxide (NO) by neutrophils (NEU) and mononuclear cells (MON) from patients with chronic granulomatous disease (CGD). Five patients with this rare disease received rHuIFN-gamma (50 micrograms/m2 of body surface, given by subcutaneous injection three times a week) for 6 months. Clinical and laboratory evaluations were performed before and after 1 and 6 months of rHuIFN-gamma therapy. Nitric oxide release by NEU and MON was assessed by the ability of these cells to inhibit thrombin-induced washed platelet aggregation. The nitrite (NO2-) and nitrate (NO3-) levels in the supernatant of cultured NEU and MON, as well as in plasma and urine (24 h diuresis), were quantified by high-performance liquid chromatography (HPLC). Conventional immunologic tests for assessing phagocyte and lymphocyte functions and humoral immunity were also performed. Therapy with rHuIFN-gamma for 6 months did not enhance NO synthesis by NEU or MON from the patients with CGD. The urinary but not plasma levels of NO2- and NO3- were elevated after rHuIFN-gamma therapy. Phagocyte and lymphocyte functions as well as humoral immunity were not affected by rHuIFN-gamma therapy. Although few patients were available for the study, we conclude that therapy with rHuIFN-gamma for 6 months did not enhance the synthesis of NO by NEU and MON in CGD patients. Whether the increased excretion of NO2- and NO3- in the urine of CGD patients after rHUIFN-gamma therapy reflects an induction of NO-synthase in cells other than leukocytes remains to be investigated.

Evaluation of a fluorochrome assay for assessing the bactericidal activity of neutrophils in human phagocyte dysfunctions.

We evaluated a method for the assessment of the phagocytic and bactericidal activity of human peripheral neutrophils against Staphylococcus aureus Cowan I, which is a modified version of the acridine orange staining technique originally described by Smith and Rommel (1977). The modification consisted of the use of free leukocyte suspensions rather than coverglass adhered leukocytes in order to avoid two main problems: the inefficient neutrophil adherence to glass that can be observed in specimens from patients with certain functional phagocyte defects, and the risk of selecting among neutrophils. An additional advantage of the modified procedure is that it permits a uniform bacteria: phagocyte ratio in different cell samples. The method was tested on 25 healthy adults and on four children with functional phagocytic defects (chronic granulomatous disease of infancy, Chediak-Higashi syndrome, and Rothmund-Thomson syndrome associated to persistent neutropenia and low chemotactic response). The neutrophils of all four patients showed a low bactericidal activity, with percent values of intracellular killed bacteria below the mean +/- 2 SD range observed in the healthy population at all incubation times tested (5, 15 and 30 min). A significant reduction in phagocytosis index and in % killed unopsonized S. aureus was observed in relation to bacteria treated with a pool of normal human serum. These results demonstrate the high sensitivity of the method, which could be used to determine intrinsic and extrinsic functional alterations in human neutrophils.

The antimeningococcal vaccine VAMENGOC B-C induced poor serum and salivary antibody response in young Brazilian children.

In 1989 about 2.3 million Brazilian children received the antimeningococcal vaccine VAMENGOC B-C (Havana, Cuba). We evaluated the serum and secretory immune response of vaccinated children by enzyme-linked immunosorbent assay with outer membrane complex antigens. Western blotting and bacterial adherence inhibition assays with human buccal epithelial cells were performed with some of the samples. Serum and salivary antibody concentrations to Neisseria meningitidis Group B of vaccinated children < 4 years old were not significantly higher than those of nonvaccinated children, as observed in convalescing patients used as positive controls. Older children (4 to 6 years old) presented a slight increase in antibody OD indexes. Sera and saliva from vaccinated children showed a weak reaction with meningococcal antigen by Western blotting and were unable to inhibit significantly the adherence of N. meningitidis B to buccal epithelial cells. These data suggest that this vaccine induced a poor serum and salivary antibody response in the population studied.

Inhibition of enteroaggregative Escherichia coli adhesion to HEp-2 cells by secretory immunoglobulin A from human colostrum.

BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is an important agent of the persistent diarrhea among low socioeconomic level children in developing countries that may be associated with chronic undernourishment. Breast-feeding is effective in protecting infants against diarrhea and other infectious diseases. The aim of the study is to verify the ability of human colostrum to inhibit aggregative adhesion of EAEC to HEp-2 cells and the presence of antibodies reactive to antigenic fractions of EAEC in colostrum samples. METHODS: Enzyme-linked immunosorbent assay, immunoblotting and adhesion assays of EAEC to HEp-2 cells were done with pooled or individual colostrum samples (n = 35). Assays were performed with a well-known EAEC strain, 044:H18 E. coli (strain 042). Colostral IgA was isolated by affinity chromatography in Sepharose anti-human alpha chain column. RESULTS: Total colostrum and isolated IgA inhibited EAEC adhesion, and this ability was associated with the presence of IgA antibodies against a 15-kDa band, compatible with the subunits of aggregative adherence fimbrial adhesin II, characteristic of the 042 strain, absent in its plasmid-cured isogenic strain, that was used as control. Individual colostrum samples also inhibited adhesion, showed variable antibody titles against EAEC antigens in enzyme-linked immunosorbent assay and recognized many antigenic fractions in immunoblotting assays, including the 15-kDa band. CONCLUSIONS: These results confirm that IgA from human colostrum inhibits adhesion of EAEC to HEp-2 cells and suggest that colostrum IgA antibodies reactive to EAEC antigens may play a role in protection of infants against diarrhea caused by these bacteria.

Functional evaluation of human neutrophils. Is the bactericidal activity correlated with nitroblue tetrazolium reduction?

The cytochemical nitroblue tetrazolium (NBT) reduction test continues to be used in clinical laboratories to detect defects in the oxidative metabolism of phagocytes. However, the specificity of the test is controversial, and it is not clear whether NBT reduction really reflects the microbicidal activity of these cells. In the present study, we evaluated the killing of Staphylococcus aureus by neutrophils from healthy adult individuals and from patients with phagocyte dysfunctions using a fluorochrome phagocytic assay, and compared the results with those obtained with a cytochemical NBT test performed simultaneously. The ability of neutrophils to reduce NBT (expressed as percent reducing neutrophils) with or without a lipopolysaccharide stimulus was not correlated with the bactericidal activity of these cells (expressed as percent killed bacteria per 100 neutrophils). The age and sex of the healthy adults did not influence the results of either assay. It seems that the superoxide anion played a small role in NBT reduction by normal neutrophils, since superoxide dismutase did not significantly inhibit this reaction. Only the absolute absence of NBT reduction reflected the low bactericidal activity of neutrophils, as seen in patients with chronic granulomatous disease (CGD). We conclude that the only clinical usefulness of the NBT test is for the screening of CGD, and that bacterial phagocytic assays are more appropriate for assessing the microbicidal function of neutrophils.

Laboratory screening for the diagnosis of children with primary immunodeficiencies.

To select simple and low-cost laboratory tests that contribute to the diagnosis of primary immunodeficiencies (PID) in children, the medical records of 98 patients with PID were analyzed. White blood cell counts, serum IgG, IgM and IgA determinations, Shick test, isohemagglutinin titers, delayed cutaneous hypersensitivity tests, nitro blue tetrazolium test and hemolytic complement (CH50) determination associated with clinical data led to the diagnosis in 95% of cases. Applying this laboratory screening to patient series studies in other countries, the diagnosis was suggested in at least 92% of the cases. This screening proposal may be used as a guideline in the standardization of complementary tests for the diagnosis of immunodeficiencies in pediatric centers of developing countries and also at the primary medical care level in developed countries.

Measurement of salivary, urinary and fecal secretory IgA levels in children with partial or total IgA deficiency.

We measured salivary, urinary and fecal secretory IgA (sIgA) levels in 11 children with total IgA deficiency and in 6 children with partial IgA deficiency using an ELISA technique. This was based on flexible microplates coated with antisecretory component (SC) and peroxidase-conjugated anti-IgA as a second antibody. Selective IgA deficiency is diagnosed as a serum IgA concentration < or = 0.05 g/l; partial IgA deficiency is diagnosed as a serum concentration of IgA > 0.05 g/l but 2 SD below normal levels. No salivary or fecal sIgA, and only low levels of urinary sIgA, were detected in the selective IgA-deficient group. The partial IgA-deficient children presented with low levels of salivary, urinary and fecal sIgA. Fecal sIgA levels correlated with salivary sIgA levels (p < 0.01) but not with urinary sIgA levels (p > 0.05) in the IgA-deficient patients. We found that all the children with partial IgA deficiency, except one, had detectable, but low values of secretory IgA. Our data suggest that these patients also have a partial mucosal IgA deficiency.

Compensatory levels of salivary IgM anti-Streptococcus mutans antibodies in IgA-deficient patients.

The protective role of salivary IgA in dental caries has not been completely demonstrated, so, in order to elucidate this point, we evaluated 15 totally and partially IgA-deficient children in terms of the following variables: dental caries indexes, bacterial plaques, number of Streptococcus mutans and Lactobacillus in the saliva, and titers of IgA, IgG and IgM anti-Streptococcus mutans antibodies in the saliva. Age-matched healthy children served as the control group. IgA-deficient children showed caries scores lower than those of the healthy children; in addition, no statistical difference was found between amount of dental plaque and numbers of the bacteria in saliva. The totally IgA-deficient children presented IgM in levels much higher than the healthy children (p < 0.05). These data could indicate a compensation for the IgA deficiency by IgM.

Application of a fluorochrome-lysostaphin assay to the detection of phagocytic and bactericidal disturbances in human neutrophils and monocytes.

The evaluation of phagocytic and microbicidal activities of the blood neutrophils has been recognized as one of the important tools for investigating phagocytic dysfunctions in patients with recurrent infections. In the present study, these activities were examined in neutrophils and monocytes from healthy adults and patients affected by primary phagocytic dysfunctions by using a modified fluorochromic microbicidal assay, discriminating simultaneously the extracellular adherence, ingestion and intracellular killing of Staphylococcus aureus Cowan I. The assay employs acridine orange staining, as described in Bellinati-Pires et al. (1989) (AO assay), but was modified by the addition of an alternative leukocyte treatment with 0.5 U/ml of lysostaphin (LS) for 5 min at 37 degrees C, after phagocytosis (AO-LS assay). The LS treatment was standardized to eliminate staphylococci adhered to the outer surface of the phagocytes without affecting the determination of intracellular live or dead bacteria, as demonstrated in normal neutrophils and monocytes. Our purpose in this study was to compare AO and AO-LS assays in order to evaluate the effect of LS on the determination of actually ingested staphylococci and to provide a means for improving the fluorochromic assay for detecting phagocytic defects, as well as bactericidal disturbances. By using the AO-LS assay, decreased ingestion of staphylococci by neutrophils in Chediak-Higashi Syndrome (CHS) was demonstrated. However, increased staphylococci adherence, as well as ingestion, was observed in neutrophils or monocytes from chronic granulomatous disease (CGD) patients, comparing AO and AO-LS assays. Bactericidal defect, which is a common feature in CHS and CGD, was detected in neutrophils or monocytes in both assays. We emphasize that such alterations were deduced by comparing the patients' results with those obtained from their respective normal controls and with the normal range of values previously established for 160 healthy adults. No alteration was observed in hyper IgE syndrome phagocytes. Despite the possible penetration of LS into the leukocytes, as stated in other studies, we concluded that a short period of phagocyte incubation with this enzyme increased the sensitivity of the fluorochromic assay to detect phagocytic defect without affecting the determination of the bactericidal activity. Moreover, comparations between AO and AO-LS assays may be important in the study of the initial pathways of staphylococci phagocyte interaction, including adherence by non-phagocytic receptors.

Phagocytic activity of human colostrum macrophages.

1. We evaluated the ability of human colostrum adhering cells to phagocytize sheep red blood cells (E) incubated with rabbit anti-E IgG antibody (A) and zymosan particles incubated with fresh human serum or with the aqueous phase of colostrum. 2. The cells were found to have considerably intense phagocytic ability, i.e., 96.8% phagocytized EA particles, 83.2% phagocytized zymosan particles opsonized with fresh human serum, and 73.3% phagocytized zymosan particles opsonized with the aqueous phase of colostrum. Thus, the aqueous phase of colostrum can opsonize zymosan particles, an activity attributed to the complement system. 3. Total hemolytic complement (CH50) and the C3 component in a pool of normal human serum were two-fold higher than in a pool of the aqueous phase of colostrum. 4. These results indicate the existence of Fc gamma and C3 receptors on the membrane of human colostrum macrophages and suggest that these cells may be biologically active.

Primary immunodeficiency diseases: a presentation of 6 cases.

The peripheral blood leukocytes of 6 children with clinical data suggestive of primary cellular immunodeficiencies were studied in an attempt to define the cellular basis of these disorders. The phenotype and function of T and B cells were investigated. According to the clinical and laboratory features, the patients were classified as one case of severe combined immunodeficiency (SCID), two of ataxia-telangiectasia (AT), one of Wiskott-Aldrich syndrome (WAS), one of DiGeorge syndrome (DSG), and one of cellular immunodeficiency (CID). The laboratory investigations together with the clinical manifestations permitted a diagnosis of primary immunodeficiency diseases.

Effect of medium- and long-chain triglycerides on human neutrophil migration.

There is some controversy concerning the effect of intravenous long-chain triglyceride (LCT) emulsions on the phagocytic system and little is known about the effect of medium-chain triglyceride (MCT)-containing emulsions. We evaluated the chemotaxis and random migration of human neutrophils from 18 healthy adults after preincubation with the following fat emulsions: LCT, MCT and a mixture of 50% MCT and 50% LCT (MCT/LCT). Leukocyte-rich plasma (4 x 10(6) cells/ml) was diluted 4:1 (v/v) with commercial fat emulsions (LCT, MCT, or MCT/LCT, 1:1) or saline and tumbled at 20 cycles/min for 30 min at 37 degrees C. The final composition of the emulsion was 20 mg/ml fat, 0.24% egg yolk lecithin, and 0.5% glycerol and the dispersion was made isotonic by adding NaCl. In a second set of experiments, the LCT and MCT concentrations were adjusted to be equimolar. Leukocyte viability was > or = 95% after exposure to the treatment with fat emulsions. For emulsions with the same weight of each fat, random migration and chemotaxis of neutrophils were unaffected by the LCT emulsion but there was a significant decrease in both chemotaxis and random migration in MCT-(79 and 74%) or MCT/LCT-treated (60 and 56%) neutrophils. Similar results were obtained when LCT and MCT were equimolar. These results demonstrate an inhibitory effect of MCT on two human neutrophil functions which may be dose dependent.

The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease.

Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis--a simple, economical and rapid method--was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.

Inherited C3 deficiency of the complement system.

1. We report a patient homozygous for C3 deficiency and several heterozygotes from the same family. Upon follow-up, the homozygote was found to suffer several severe bacterial infections, whereas all the heterozygotes were clinically healthy. 2. C3 was undetectable in the homozygous patient, CH50 was very low and factor I was present. Serum capacity to generate chemoattractant stimuli for peripheral leucocytes was similar to that of normal adults as was also observed for one of the heterozygotes. Serum capacity to opsonize yeast was reduced in the presence of autologous and homologous (normal adult) cells. The CH50 levels of heterozygous patients were within the lower range of normality. 3. The parental consanguinity and the homozygosis state observed here are classical signs of recessive autosomal inheritance. However, the lower or below normal C3 levels detected in parents and relatives point to a co-dominant inheritance of gene S with respect to the "null" gene. 4. C3 polymorphism presented a predominantly "slow" pattern in most family members, which, together with the low C3 levels, indicates the expression of S-allotypes.

Transfer of IgG subclasses across placenta in term and preterm newborns.

In order to study placental transfer of IgG subclasses, paired blood samples were collected from mothers and umbilical cord of preterm (N = 69) and full-term (N = 68) newborns. The full-term group was further divided into 3 subgroups: appropriate for gestational age (AGA, N = 43), large for gestational age (LGA, N = 13) and small for gestational age (SGA, N = 12), according to birth weight. IgG subclasses (IgG1, IgG2, IgG3 and IgG4) were measured by the single radial immunodiffusion technique using monoclonal antibodies. IgG1 and IgG3 newborn subclass concentrations (10.17 and 0.57 g/l, respectively) increased with increasing gestational age and reached maternal levels (IgG1 = 8.86; IgG3 = 0.67 g/l) during the 37th week of pregnancy. Low levels of these subclasses were found in premature newborns. IgG2 from newborns were always lower than maternal levels (P < 0.05). LGA and SGA newborns had equivalent levels of IgG1 and IgG2 compared with AGA. SGA newborns had higher levels of IgG3 and lower levels of IgG4 than LGA and AGA newborns.

An in vitro model to study the effect of antibodies and cell receptor analogues on meningococcal adherence to human oroepithelial cells.

We have investigated different experimental schedules to achieve adherence of Neisseria meningitidis group B to cultured and buccal epithelial cells (BEC) and the effect of antibodies and receptor analogues on bacterial adherence. No adherence of meningococcus was observed when HeLa, HEp-2 or KB cells were used, but high rates of adherence to BEC occurred. The effect of antibodies on bacterial adherence was studied in assays carried out in the presence of saliva and serum collected from convalescing children with meningococcal meningitis and children vaccinated with VAMENGOC B-C. Both saliva and serum from the convalescent patients inhibited the adherence of meningococci, but saliva and serum from vaccinated children did not, corroborating our previous data of a poor antibody response induced by this vaccine. Human colostrum did not affect meningococcal adherence despite the presence of antibodies to N. meningitidis detected by ELISA. Inhibition of adherence by sera from an immunized horse, rabbits and mice, as well as by cell receptor analogues (outer-membrane complex and purified polysaccharide C), was observed. Our results show that up to now BEC continue to be the best cells to study meningococcal adherence and the effect of adherence inhibitors.