In-vivo immune responses of breast- and bottle-fed infants to tetanus toxoid antigen and to normal gut flora

The effects of breast- and bottle-feeding on serum immunoglobulin levels and specific antibody responses have been examined in 30 infants on five occasions from 6 days until 9 months of age. No significant differences were found on any sample occasion between the two feeding groups in total immunoglobulin levels of G, M and A classes or in class-specific antibody responses to tetanus toxoid vaccine. This suggests that the capacity of the two groups to make serum antibodies develops similarly. Concentrations of antibodies to commensal Escherichia coli 'O' lipopolysaccharide antigens, however, were significantly greater in the bottle-fed group, and it is suggested that this difference is due to an increase in the exposure of the systemic immune system to these gut antigens in the bottle-fed infants. There are several possible explanations for this increased exposure and the resulting effects on the infants' immune system. These experiments also illustrate a possible role of breast milk in stimulating the immune system.     

ANTIBODY PRODUCTION BY THE MAMMARY GLAND IN MOTHERS AFTER ARTIFICIAL ORAL COLONISATION OF THEIR INFANTS WITH A NON-PATHOGENIC STRAIN E. COLI 083

Abstract. Twenty five breast-fed and 25 formula-fed infants were colonised by oral administration of a living suspension of E. coli 083. Twenty breast-fed and 13 formula-fed infants were followed as controls. Specific antibody titres in serum, stool filtrates and milk, and secretory IgA levels in stool filtrates and milk were determined in samples taken fortnightly from birth until 20 weeks of age. The haemagglutinating antibody in serum and milk increased in the colonised groups, but in stool filtrates an inhibitory effect of breast-milk was demonstrated. Secretory IgA levels in stool filtrates were significantly higher in colonised infants and breast-fed controls than in bottle-fed infants during the period of breast feeding. Then levels in the colonised groups remained high, but in breast-fed controls they decreased to values found in bottle-fed controls. Artificial colonisation evoked local antibody and secretory IgA responses in the intestine, as well as an antibody response in the mother's mammary gland. The possible protective effect of those responses is discussed.     

Antibody production by the mammary gland in mothers after artificial oral colonisation of their infants with a non-pathogenic strain E. coli 083.

Twenty five breast-fed and 25 formula-fed infants were colonised by oral administration of a living suspension of E. coli 083. Twenty breast-fed and 13 formula-fed infants were followed as controls. Specific antibody titres in serum, stool filtrates and milk, and secretory IgA levels in stool filtrates and milk were determined in samples taken fortnightly from birth until 20 weeks of age. The haemagglutinating antibody in serum and milk increased in the colonised groups, but in stool filtrates an inhibitory effect of breast-milk was demonstrated. Secretory IgA levels in stool filtrates were significantly higher in colonised infants and breast-fed controls than in bottle-fed infants during the period of breast feeding. Then levels in the colonised groups remained high, but in breast-fed controls they decreased to values found in bottle-fed controls. Artificial colonisation evoked local antibody and secretory IgA responses in the intestine, as well as an antibody response in the mother's mammary gland. The possible protective effect of those responses is discussed.     

Serum antibody levels and avidities to Escherichia coli O antigens and poliovirus type 1 antigen are increased in children treated for malignant disease.

BACKGROUND: Treatment of malignant disease in children is often associated with low serum immunoglobulin and reduced specific antibody levels. The aim of this study was to investigate if the functional affinity of specific antibodies in serum and saliva is reduced as well and to evaluate if antigenic exposure or treatment duration affects this antibody avidity. PROCEDURE: Serum samples were obtained from 45 children and salivary specimens from 30 children with malignant disease. The children were tested either prior to, during, or after chemotherapy. Levels of antibody to E. coli O and to poliovirus type 1 antigens were determined using an ELISA and isotype-specific relative antibody avidity was measured using thiocyanate to elute antibodies from solid-phase immobilized antigens. RESULTS: Children with malignant disease had higher levels and relative avidity indexes of serum antibodies to both antigens as compared to controls. The duration of treatment and type of malignant disease were unrelated to these parameters. In saliva, the level of antibodies to E. coli O antigens, but not to poliovirus type 1 antigen, increased during treatment. CONCLUSIONS: Both the amount and avidity of serum antibodies to these antigens are increased in children with malignant disease. This may be due to a dysregulation of the immune system caused by the malignancy and seems not to be dependent on exposure. In contrast, the avidity and levels of these antibodies in saliva seem to correlate with the presence of antigenic exposure.     

Antibody levels to Bordetella pertussis in 10-yr-old children with atopy and atopic asthma

Suboptimal immune responses to vaccination have been suggested among atopic infants. The aim of this study was to assess the influence of atopy and atopic asthma on the humoral response to Bordetella pertussis vaccination. Immunoglobulin (Ig)G and IgA specific antibodies were measured by enzyme linked-immunosorbent assay in 102, 10-yr-old atopic children (66 of them also being asthmatics) and compared with 76 non-atopic and 53 non-atopic non-asthmatic controls of similar age. The levels of antibodies and the percentage of positives to B. pertussis were comparable in all groups. Children with a very high total serum immunoglobulin (Ig)E (Percentile (Pct) > 90th) showed higher (p = 0.01) IgG pertussis antibodies than children with very low serum IgE (Pct < 10th). In conclusion, we found normal pertussis antibody levels in atopic and in atopic asthmatic children in late childhood, thus overriding any possible suboptimal response during infancy.     

Development of immunoglobulin G and immunoglobulin E antibodies to cow's milk proteins and ovalbumin after a temporary neonatal exposure to hydrolyzed and whole cow's milk proteins

The ingestion of food antigens usually results in the induction of oral tolerance, but the clinical and immunologic consequences of brief exposure to cow's milk proteins during the neonatal period are not well‐documented. The aim of this work was to study immunoglobulin (Ig)E and IgG responses to cow's milk proteins and ovalbumin after exposure during the first three days of life in infants who were otherwise exclusively breast‐fed. A group of 129 infants was randomly assigned at birth to one of three feeding regimens: human milk (HM), cow's milk formula (CMF), or a casein hydrolysate formula (CHF), during the first three days of life. They were then all exclusively breast‐fed for a varying period of time and followed for two years. Serum IgG and IgE antibodies to cow's milk proteins and ovalbumin (OVA) were analyzed in blood samples obtained at birth, at 4 days and at 2, 4, 8, 12 and 24 months of age. The levels of IgG antibodies to β‐lactoglobulin (IgG‐BLG) and bovine serum albumin (IgG‐BSA) were higher in the CMF and the HM groups than in the CHF group for up to two years. This was particularly obvious for IgG‐BLG in infants who started weaning before two months. The levels of IgG antibodies to casein (IgG‐CAS) were higher in the CMF goup, as compared with the CHF group at 8 and 12 months. The levels of IgG antibodies to OVA were similar in all three feeding groups. The levels of IgE antibodies to CAS or OVA were similar in the three feeding groups. Exposure to cow's milk during the first three days of life stimulated IgG antibody production to cow's milk proteins and this was still obvious at 2 years of age, while feeding with a casein hydrolysate during the first three days of life was associated with low levels of IgG antibodies to cow's milk proteins.     

Food protein-induced enterocolitis: altered antibody response to ingested antigen

To evaluate the role of immunologic mechanisms in one specific syndrome of food intolerance in infants, food protein-induced enterocolitis, we measured class-specific serum antibodies to three food proteins, ovalbumin, soy, and cow milk, prior to diagnostic food challenges in 18 infants suspected to have this syndrome. Infants with positive challenge reactions to egg, soy, or cow milk had 5-10 times higher levels of IgA antibody directed against that food than did the infants with negative challenges. Levels of IgG antibody to soy and egg were also significantly higher (greater than 10-fold) in infants with positive challenge responses. There was no significant difference in levels of IgM food antibodies between the two groups. IgA anti-soy antibody levels rose in all 12 infants tested 2-10 weeks after a single soy feeding (challenge). However, IgM anti-soy antibody increased in the five infants who had a negative response to the challenge feeding and decreased in those seven with a positive response. The difference between the two groups was statistically significant (P less than 0.01). Some correlation existed (r = -0.68) between the increase in IgA anti-soy antibody and the decrease in IgM anti-soy antibody for infants with positive soy challenges. Although a pathogenic role for these antibodies is not proven, the findings suggest an altered immunologic response to ingestion of food antigens in infants with food protein-induced enterocolitis.     

A prospective study of humoral immune responses to cow milk antigens in the first year of life

Previous studies have shown that in cow milk allergy the specific immune response to dietary cow milk antigens is deficient. This study aimed at delineating the development of humoral immune response to cow milk antigens in healthy infants. Twenty-five healthy newborns were enrolled, and seen at scheduled visits at the ages of three, six and eleven months, and they formed two groups: those breastfed and those fed adapted cow milk formulae. The local immune response in the gut was approximated using the ELISPOT assay of circulating antibody secreting cells. At the age of three months, in the formula fed group, cells secreting specific IgA to cow milk antigens were detected despite low levels of IgA serum antibodies. The total number of IgA secreting cells increased with age (p = 0.001). The milk in the infant diet directly influenced this development so that the age related increase was significantly greater in the formula fed group (p = 0.04). The results indicate that diet has a significant effect on the developing immune system, and that healthy infants are able to respond in an antigen specific fashion to dietary antigens, which may be central in attaining clinical tolerance of such antigens.     

Titres of specific antibodies to poliovirus type 3 and tetanus toxoid in saliva and serum of children with recurrent upper respiratory tract infections

A study of antibody levels (in saliva and blood) against common vaccine antigens was performed in a population of 32 children suffering from recurrent upper respiratory tract infections (URTI). None of the patients had primary or secondary immunodeficiency syndromes or other known predisposing factors for respiratory diseases. Titres of the isotype-specific antibodies immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin G (IgG) against two vaccine antigens – poliovirus type 3 (P3) and tetanus toxoid (TT), a viral antigen and a bacterial antigen, respectively – were measured in unstimulated saliva and serum, both in patients and in 24 healthy children (controls), by using a standard enzyme-linked immunosorbent assay (ELISA). In addition, levels of total IgA and avidity of IgA antibodies to both P3 and TT in saliva were evaluated. No difference was found between patients and controls as to levels of total IgA, or specific IgA and IgM antibodies against both P3 and TT in saliva. Furthermore, the avidity of salivary IgA antibodies against the two antigens did not differ between the two populations. However, the average concentrations of saliva-specific IgG antibodies to both the viral and the bacterial antigen were significantly lower (p < 0.01 for P3 and p < 0.05 for TT, respectively) in saliva of children with recurrent URTI, whereas no difference was found in serum for any immunoglobulin isotype determined compared with healthy individuals. The results of the present study provide suggestive evidence for the existence of subtle IgG-restricted defects in antibody responses at the mucosal level, but not at the serum level, in some children with undue susceptibility to URTI.     

Mucosal immune responses to meningococcal conjugate polysaccharide vaccines in infants

BACKGROUND: Serogroup C meningococcal conjugate polysaccharide vaccines have been reported to induce significant serum IgG antibodies and immunologic memory in infants. Because meningococcus is a mucosal pathogen colonizing the nasopharynx, local mucosal immune responses may play an important role in host defense against infection and carriage. We have investigated the mucosal IgA and IgG antibody responses to two meningococcal C conjugate vaccines in the saliva of healthy infants. METHODS: Specific salivary IgA and IgG antibodies to two meningococcal C polysaccharide conjugate vaccines (Menjugate from Chiron Corp., n = 46; and Meningitec from Wyeth Lederle, n = 54) were investigated by immunoassay in infants after parenteral vaccinations at the ages of 2, 3 and 4 months. Unstimulated saliva samples were collected immediately before the first immunization and 1 month after the third immunizations. Forty healthy infants receiving the same routine vaccines but no meningococcal C vaccine were recruited as controls. RESULTS: There were significant increases in meningococcal C polysaccharide-specific IgG antibody concentrations postvaccination compared with prevaccination concentrations in both vaccinated groups (both P < 0.001), but no change in the control group. There were no significant increases in specific IgA postvaccination geometric mean concentrations in either the vaccine or the control groups. The number of IgA positives postvaccination increased slightly in the Wyeth vaccine group vs. controls (P < 0.05). CONCLUSIONS: Significant salivary IgG antibodies to meningococcal C polysaccharide were observed after parenteral immunization with two meningococcal C conjugate vaccines, whereas there was no significant increase in specific IgA antibody levels for these two vaccines.     

Antibody Responses to Parenteral and Oral Vaccines Are Impaired by Conventional and Low Protein Formulas as Compared to Breast-feeding

ABSTRACT. The vaccine response to poliovirus, diphtheria and tetanus toxoids in relation to protein intake was studied in infants, either breast-fed or given low (1.1 g/100 ml) or conventional (1.5 g/100 ml) protein formula. Serum, salivá and faeces antibodies were measured by the enzyme-linked immunosorbent assay. Neutralizing poliovirus antibodies were determined. The serum, saliva and faeces antibody responses in the two formula-fed groups of infants did not differ significantly, but for the low protein formula group which had significantly higher serum neutralizing titres to poliovirus after the second vaccine dose than the conventional formula group. However, the breast-fed group had significantly higher antibody levels than the two formula-fed groups together: serum IgG to diphtheria toxoid (p<0.01) and serum neutralization of poliovirus (p<0.001) at 21-40 months of age, saliva secretory IgA to tetanus (p<0.01), diphtheria toxoid (p<0.01) and poliovirus (p<0.05), as well as faecal IgM to tetanus toxoid (p<0.05) and poliovirus (p <0.01 and p <0.05) at 3 and 4 months of age. Breast-fed infants thus showed better serum and secretory responses to peroral and parenteral vaccines than the formula-fed, whether with a conventional or low protein content.     

ESCHERICHIA COLI O ANTIBODY CONTENT IN MILK FROM HEALTHY SWEDISH MOTHERS AND MOTHERS FROM A VERY LOW SOCIO-ECONOMIC GROUP OF A DEVELOPING COUNTRY

Abstract. The antibody content of milk from healthy Swedish mothers was compared with that of milk from mothers of a very low socio-economic group in a developing country. Antibodies of various immunoglobulin classes against E. coti O antigens were determined with the enzyme-linked immunosorbent assay (ELISA). The milk antibodies which mainly belonged to the secretory IgA class appeared in similar concentrations in milk from the two groups using E. coli antigens of Swedish as well as Pakistani origin. The secretory IgA antibodies could be demonstrated in the stool of the breast-fed infants of the undernourished mothers. Also the concentration of serum IgG and IgA antibodies to E. coli O antigens were similar in the Pakistani and Swedish mothers. The serum IgM antibody levels of the Pakistani mothers were higher, however, presumably due to a higher frequency of infections. It was noted that the milk production decreased considerably upon the hospitalization of the healthy and well-to-do Swedish mothers. The small milk volumes of the undernourished Pakistani mothers suggest that the lactation failure observed was mainly due to inadequate milk flow and not to decreased milk quality. The results indicate the necessity of studying the nutritional, psychological and social factors responsible for low milk yield and add yet another reason to stimulate prolonged breastfeeding.     

Serum bile acids and their conjugates in breast-fed infants with prolonged jaundice

Serum bile acids and their conjugates were analysed in 20 breast-fed infants with prolonged jaundice. The mean total bile acid levels in serum were increased in the breast-fed infants with jaundice, as compared with those in either breastor bottle-fed infants without jaundice. However, there were no significant differences between the groups. All the breast-fed infants examined, regardless of association with jaundice, had a bile acid pattern dominated by taurine conjugates (the ratio of glycine- to taurine-conjugated bile acid, G/T ratio, less than 1.00). In contrast, the bottle-fed infants without jaundice had a pattern dominated by glycine conjugates (G/T ratio, more than 1.00). Among the breast-fed infants with jaundice, the mean G/T ratio in those who had serum bilirubin levels over 10 mg/100 ml was significantly lower than that in those who had serum bilirubin levels of less than 10 mg/100 ml. The altered bile acid metabolism might be associated with the pathology of breast milk jaundice.Abbreviation LP-X lipoprotein-X     

Antibody responses to parenteral and oral vaccines are impaired by conventional and low protein formulas as compared to breast-feeding

The vaccine response to poliovirus, diphtheria and tetanus toxoids in relation to protein intake was studied in infants, either breast-fed or given low (1.1 g/100 ml) or conventional (1.5 g/100 ml) protein formula. Serum, saliva and faeces antibodies were measured by the enzyme-linked immunosorbent assay. Neutralizing poliovirus antibodies were determined. The serum, saliva and faeces antibody responses in the two formula-fed groups of infants did not differ significantly, but for the low protein formula group which had significantly higher serum neutralizing titres to poliovirus after the second vaccine dose than the conventional formula group. However, the breast-fed group had significantly higher antibody levels than the two formula-fed groups together: serum IgG to diphtheria toxoid (p less than 0.01) and serum neutralization of poliovirus (p less than 0.001) at 21-40 months of age, saliva secretory IgA to tetanus (p less than 0.01), diphtheria toxoid (p less than 0.01) and poliovirus (p less than 0.05), as well as faecal IgM to tetanus toxoid (p less than 0.05) and poliovirus (p less than 0.01 and p less than 0.05) at 3 and 4 months of age. Breast-fed infants thus showed better serum and secretory responses to peroral and parenteral vaccines than the formula-fed, whether with a conventional or low protein content.     

Dietary factors influencing levels of food antibodies and antigens in breast milk

The effect of a milk-free diet during late pregnancy and lactation, on levels of cows'milk specific antibodies and antigens in breast milk, was examined. Women with an allergic condition (defined as atopic) were randomly allocated to a milk-free diet ( n = 10) or an unrestricted diet ( n = 12). Twelve non-atopic women followed an unrestricted diet. A significant reduction ( p < 0.001) in β-lactoglobulin-specific immunoglobulin A and α-casein-specific immunoglobulin A levels was observed over the first 5 days in all groups. The mean level of β-lactoglobulin antigens in breast milk from women who adhered strictly to the milk-free diet was significantly lower than the levels of the atopic group on the unrestricted diet ( p < 0.02). The allergy incidence in the infants born in the atopic diet group was significantly lower compared with that of the atopic group on the unrestricted diet.     

Oral rotavirus vaccination in breast- and bottle-fed infants aged 6 to 12 months

Oral bovine rotavirus vaccine RIT 4237 was tested at two dose levels in 217 children between 6 and 12 months of age who received either breast milk or bottle milk (cow's milk or infant formula) before vaccination. Of the children 65% were initially seronegative for rotavirus ELISA IgG and IgM antibody. The full vaccine dose (10(8.3) 50% tissue culture infective doses, TCID50) induced seroconversion rates 81% and 86% and booster response rates 69% and 64% in breast and bottle-fed children, respectively. There was no difference in the vaccine response of infants receiving cow's milk or infant formula either. It is concluded that the RIT 4237 rotavirus vaccine at the dose level 10(8.3) TCID50 gives a satisfactory response in both breast- and bottle-fed children in this age group, but multiple vaccinations may be needed for maximal efficacy.     

ANTIBODIES AGAINST ESCHERICHIA COLI CAPSULAR (K) ANTIGENS IN HUMAN MILK AND SERUM

ABSTRACT. Specimens of milk, serum and faeces were collected from 23 healthy women, as well as faeces from their newborn infants, at several occasions from one to nine weeks after delivery. The predominating faecal E. coli strains were typed with regard to five different K antigens. The most frequently occurring was K1, followed by K3 and K13. E. coli with the same K antigen was found in only five of the mother-infant pairs, while ten pairs never had any strain with a common K antigen during the observation period. Milk and serum antibodies were regularly found against a variety of E. coli K antigens using the enzyme-linked immunosorbent assay. In milk the dominating immunoglobulin class of anti-K antibodies was secretory IgA, with a high ratio of milk/serum antibody, suggesting a local production in the mammary gland. There were no significant differences of K antibody levels in serum or milk, whether or not the particular K strain was present in the mother's gut flora. The presence of specific K antibodies in serum or milk of the mothers did not prevent colonization of the gut of the infants with E. coli strains carrying the corresponding K antigen.     

Effect of breast-feeding on seroresponse of infants to oral poliovirus vaccination.

Three hundred Indian infants between 6 and 51 weeks of age were divided into six groups and given three doses of trivalent oral polio vaccine (OPV) of known adequate potency. One group was on unrestricted breast-feeding with mandatory breast-feed during the interval between 30 minutes before and 15 minutes after each dose of OPV. In four groups of infants breast-feeding was withheld for three, four, five, and six hours both before and after each dose of OPV. The sixth group was bottle-fed. Samples of blood were collected from all infants before vaccination and from 227 infants further samples were collected four weeks after the first and/or third doses of OPV. Antibody responses to poliovirus types 1, 2, and 3 were determined following one dose and three doses of OPV, and the rates of response were found to be approximately equal in all groups of breast-fed infants irrespective of their feeding schedules, as well as in bottle-fed infants. Thus breast-feeding is shown to have no inhibitory effect on antibody response of infants beyond the newborn period to OPV.     

The effect of breast-feeding on proliferation by infant lymphocytes in vitro

The effect of breast-feeding on the development of lymphocyte responsiveness in infants has been studied. Peripheral blood mononuclear cells from 15 breast- and 15 bottle-fed infants were obtained sequentially between 6 days and 9 months of age. A number of agents were used to stimulate the cells in vitro and the resulting proliferative responses were compared between the two feeding groups. A hanging drop microculture system using serum-free medium, enabled spontaneous proliferation and proliferative responses to several stimuli (T and B cell mitogens, allogeneic lymphocytes, and antigen) to be studied at a range of cell concentrations and days of culture. Significant age-related differences were found between the responses of cells from the two feeding groups. Spontaneous proliferation and proliferative responses to the T cell mitogen phytohaemagglutinin and the antigen tetanus toxoid were significantly greater in the breast-fed group at the two earliest ages studied (6 days and 6 wk). Responses to mitogens which predominantly affect B cells, such as pokeweed mitogen and Staphylococcus aureus (Cowan), were similar in both feeding groups at this age. In contrast, from 3 to 9 months of age, responses of cells from bottle-fed infants were significantly greater to all stimuli than responses from breast-fed infants. One possible explanation for the higher level of proliferation by cells from newborn breast-fed infants, is that these infants may absorb the cell-growth factors and lymphokines known to be present in human colostrum and milk. These factors may stimulate T cells and/or their precursors in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)     

Impact of treatment reduction for childhood acute lymphoblastic leukemia on serum immunoglobulins and antibodies against vaccine-preventable diseases

Background

The consequences of current intensive chemotherapy for childhood acute lymphoblastic leukemia (ALL) for immune defense are a matter of concern. The purpose of this study was to examine the effect of reduced compared with intensive (conventional) ALL chemotherapy on serum immunoglobulin levels and specific antibody concentrations against vaccine‐preventable diseases.

Procedure

Patients treated according to Dutch Childhood Oncology Group ALL 10 protocol were stratified by minimal residual disease to receive reduced (standard risk; SR) or intensive (medium risk; MR) intensification/maintenance treatment. Between November 2004 and July 2009 we compared serum immunoglobulins of 110 patients and specific antibodies against diphtheria toxin, tetanus toxin, and Bordetella pertussis antigens of 41 patients of SR and MR groups during chemotherapy.

Results

Immunoglobulin levels showed significantly different patterns between the SR and MR groups. In the MR group IgG, IgA, and IgM levels decreased towards the end of intensive treatment; in the SR group IgG levels increased while IgA and IgM stabilized. In both groups IgM and IgG levels were most affected. Specific antibody levels against vaccine‐preventable diseases decreased in both groups, but more profound in MR group.

Conclusions

Although reduced chemotherapy is beneficial for immunoglobulin level recovery and might prevent susceptibility for infections, specific antibodies remain decreased. Pediatr Blood Cancer 2012; 58: 701–707. © 2011 Wiley Periodicals, Inc.