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Inhibition by colostrum of the responses of peripheral blood mononuclear cells to mitogens.

This study investigated the inhibition by colostrum of the uptake of tritiated thymidine by both autologous and heterologous peripheral blood mononuclear cells (PBMC) stimulated with the mitogens phytohaemagglutinin (PHA), pokeweed mitogen (PWM) or Concanavalin A (Con A). This inhibition was not the result of an alteration in the dose response or kinetics of mitogenesis by the colostrum. The inhibitory factor in the colostrum was neither dialysable nor lymphocytotoxic. Lymphocytes which were briefly pre-treated with colostrum, then washed and cultured, also incorporated much less labelled thymidine than control cells. The inhibitory factor was not adsorbed from colostrum by mononuclear cells. We conclude that colostrum contains a cytostatic factor which inhibits mitogenesis in PBMC.

Colostrum↗

Use of the 'nutriceutical', bovine colostrum, for the treatment of distal colitis: results from an initial study.

BACKGROUND: Bovine colostrum is a rich source of nutrients, antibodies and growth factors. AIM: To examine the efficacy of colostrum enemas in the treatment of distal colitis using a randomized, double-blind, controlled protocol. METHODS: Fourteen patients (eight female), with a mean age of 45 years (range, 16-75 years) and mild to moderately severe distal colitis (Powell-Tuck scoring system), received colostrum enema (100 mL of 10% solution) or placebo (albumin solution) b.d. for 4 weeks. Both groups also received mesalazine (1.6 g/day) or, if already taking it, had a dose increment of 1.6 g/day. Disease activity was documented at 0, 2 and 4 weeks. RESULTS: After 4 weeks, the colostrum group showed a mean reduction in symptom score of - 2.9 (95% confidence interval (CI), - 5.4 to - 0.3), whereas the placebo group showed a mean response of + 0.5 (95% CI, - 2.4 to +3.4). The histological score improved in five of the eight patients in the colostrum group (mean response, - 0.9; 95% CI, - 1.69 to - 0.03), whereas the histological scores only improved in two of the six patients in the placebo group (mean response, 0.2; 95% CI, - 2.4 to +2.6). CONCLUSIONS: Bovine colostrum enema shows potential as a novel therapy for left-sided colitis with additional benefits over using mesalazine alone. Further studies appear to be warranted.

Adolescent↗

Detection and characterization of antibodies specific to food antigens (gliadin, ovalbumin and beta-lactoglobulin) in human serum, saliva, colostrum and milk.

Antibodies against food antigens are usually produced in healthy people. This humoral response can be detected both in serum and secretions. The characterization of this response can be useful for a better understanding of food-related immunological alterations. In this study, IgA and IgG antibodies specific to ovalbumin, beta-lactoglobulin or gliadin were measured in serum, saliva, colostrum and milk from 40 healthy breast-feeding women. Specific IgA and IgG to the three antigens were measured by indirect ELISA. Specific IgG levels were highest in serum and very low in the other biological fluids. No correlation between the IgG specific to the different antigens was found. Specific IgA reactivity was found in all the samples analysed. Levels observed were higher in colostrum and milk than in serum and saliva. In spite of being three different unrelated food antigens, a correlation between the levels of specific IgA was found in saliva, colostrum and milk samples of all subjects studied. The specificity of IgA anti-gliadin antibodies from serum, saliva and colostrum was analysed by immunoblotting of SDS-PAGE-separated wheat proteins. Each sample presented a unique pattern of recognition. No common pattern of recognition was found either among the same biological fluids of the different subjects tested, or among the different samples--either serum, colostrum or saliva--of the same individual. Different degrees of specificity to wheat proteins among IgA from colostrum, saliva or serum were observed, suggesting that the local IgA-producing populations are functionally different in the different tissues of the organism.

Antibodies↗

Bovine colostrum supports the serum-free proliferation of epithelial cells but not of fibroblasts in long-term culture.

Medium lacking serum but supplemented with milk will support the growth of sparse cells in culture. Milk obtained within 8 h after the birth of a calf (day 1 colostrum) is the most effective in supporting proliferation. In mixed cultures of early-passage bovine embryonic kidney (BEK) or early-passage calf kidney (CK) cells, both epithelial cells and fibroblasts grow in Dulbecco's modified eagle's medium (DMEM) supplemented with serum. However, only cells that appear to be epithelial-like grow in DMEM supplemented with colostrum. Sparse cultures of early-passage human and rat fibroblasts that grow readily in DMEM supplemented with serum do not grow in DMEM supplemented with colostrum. Canine kidney epithelial cells (MDCK), when plated sparsely, grow exponentially in DMEM supplemented with day 1 bovine colostrum. The generation time is 26 h, the same growth rate as in DMEM supplemented with calf serum. The MDCK cells can be subcultured and regrown to confluence repeatedly in colostrum-supplemented DMEM. Growth in DMEM supplemented with colostrum does not alter the morphological characteristics of the MDCK cells, which are polygonal, contain microvilli at the apical surface, and are connected by tight junctions and desmosomes. MDCK cells do not proliferate in DMEM supplemented with milk obtained 1 wk after the birth of a calf.

Animals↗

Detection of HIV-1 genome in leukocytes of human colostrum from anti-HIV-1 seropositive mothers.

In order to obtain more information about the presence of HIV-1 in mononuclear cells of colostrum, research was carried out on both the HIV-1 genome in the cellular fraction of colostrum and the viral antibody in cell-free colostrum of eight anti-HIV-1 seropositive asymptomatic mothers. In five cases cell fractions of the colostrum harbored HIV-1 genome by DNA-DNA and DNA-RNA in situ hybridization, whereas viral antibody were detected in all cell-free colostrum specimens. The data confirms the colostrum as a possible route of HIV-1 infection.

Adult↗

Porcine colostrum and milk stimulate visceral organ and skeletal muscle protein synthesis in neonatal piglets.

Our objective was to determine the relative contributions of protein synthesis and protein absorption in the rapid accretion of gastrointestinal protein in suckling piglets during the early neonatal period. We measured the rates of tissue protein synthesis using a flooding dose of L-[4-3H]phenylalanine in various visceral and peripheral tissues of neonatal piglets fed water, mature milk or colostrum for 6 h. The jejunal and ileal protein synthesis rates in piglets fed either colostrum or milk were three- to fourfold higher than in piglets fed water. The increased jejunal and ileal protein synthesis could not, however, account for the differences in protein mass between the colostrum-fed and water-fed groups. The relative abundance of IgG, a major porcine colostral protein, in jejunal tissue was markedly higher in piglets fed colostrum than in piglets fed either milk or water. The fractional protein synthesis rates in liver, kidney, spleen and skeletal muscle and the absolute protein synthesis rates in liver and spleen were also greater in piglets fed colostrum than in those fed milk or water. Increased endogenous protein synthesis made only a minor contribution to the increased intestinal protein accretion in neonatal piglets fed colostrum. A much larger proportion of this increase seemed to be a result of absorption and retention of ingested immunoglobulins.

Animals↗

Treatment with bovine hyperimmune colostrum of cryptosporidial diarrhea in AIDS patients.

Cryptosporidium parvum may cause severe, debilitating diarrhea in patients with AIDS. Recent anecdotal reports have suggested that hyperimmune bovine colostrum may be effective. We conducted a double-blind, controlled pilot study of hyperimmune bovine colostrum for diarrhea due to cryptosporidiosis in five AIDS patients. The patients were randomized to receive either hyperimmune or control colostrum by continuous nasogastric infusion for 10 days. All stools were collected, graded, and weighed, and the concentration of oocysts excreted was determined daily. One of the three patients treated with hyperimmune colostrum had a reduction in diarrhea and in the concentration of oocysts excreted. A second treated patient had a modest decrease in the concentration of oocysts excreted. Two patients who received control colostrum also had decreases in the volume of diarrhea but no change in the concentration of oocysts excreted. We conclude that hyperimmune colostrum with high titers of specific anti-Cryptosporidium antibody could be effective in treating patients with cryptosporidiosis. However, more studies of cow colostral immunoglobulin need to be performed so that the efficacy of this treatment can be assessed more thoroughly.

Acquired Immunodeficiency Syndrome↗

Human colostrum and serum contain antibodies reactive to the intimin-binding region of the enteropathogenic Escherichia coli translocated intimin receptor.

BACKGROUND: In Brazil, enteropathogenic Escherichia coli (EPEC) diarrhoea is endemic in young infants. A characteristic feature of EPEC adhesion to host cells is intimate attachment leading to the formation of distinctive "attaching and effacing" (A/E) lesions on mammalian cells. Two genes directly involved in intimate adhesion, eae and tir, encode the adhesion molecule intimin and its translocated receptor Tir, respectively. The intimin-binding domain of Tir was recently mapped to the middle part of the polypeptide (Tir-M), and the amino (Tir-N) and carboxy (Tir-C) termini were found to be located within infected host cells. Recently, it was shown that colostrum samples from mothers living in Sao Paulo contain IgA-class antibodies reactive with a number of proteins associated with EPEC virulence. It has also been shown that patients infected with verocytotoxin-producing E. coli O157 can produce antibodies to Tir. In the current study antibody responses to the different Tir domains were analyzed in sera and colostrum samples collected in an EPEC-endemic area of Brazil. METHODS: Recombinant Tir, Tir-N, Tir-M, and Tir-C were expressed as His-tagged protein in E. coli BL21a and purified on nickel columns. Western blot analysis was used to investigate colostrum IgA- and serum IgG-class antibodies reactive with the Tir fragments. RESULTS: Anti-Tir IgG antibodies were detected in the serum of children, with (63%) or without (50%) diarrhoea. Anti-Tir IgA-class antibodies were detected in all the colostrum pools tested. With the use of both serum IgG- and colostrum IgA-class antibodies, an immunodominant domain of the Tir-polypeptide, Tir M, was identified. CONCLUSION: The intimin-binding region of Tir (Tir-M) is the immunodominant region of the polypeptide in humans. Both serum IgG-class and colostrum IgA-class antibodies reacted predominantly with the Tir-M domain.

Adhesins, Bacterial↗

Dose effects of oral bovine colostrum on physical work capacity in cyclists.

PURPOSE: There is interest in the potential long-term use of dietary supplementation with bovine colostrum to enhance exercise performance. The purpose of the present study was to determine the dose effects of bovine colostrum on cycling performance. METHODS: Forty-two competitive cyclists were randomly divided into three groups and required to consume either 20 g/d bovine colostrum + 40 g whey protein concentrate (wpc), 60 g of bovine colostrum, or 60 g of wpc (placebo). Two measures were used to assess performance before (pre-) and after (post-) an 8-wk supplementation period. The first measure required subjects to complete two VO2max tests separated by 20 min with the amount of work completed in the second test used to evaluate performance. The second performance measure was the time to complete a work-based time trial following a 2-h cycle at 65% VO2max. Subjects were required to maintain their regular training and keep a food and training diary over the study period. RESULTS: After supplementation, the performance enhancement in Measure One was not statistically significantly different in the colostrum groups compared to the placebo group (placebo = 3.4%, 20 g = 4.0%, 60 g = 3.9%; 95% confidence interval (CI) for differences, +/-1.8%, P > 0.05). In performance Measure Two subjects in the 20 g and 60 g groups completed the time trial significantly (P < 0.05) faster post supplement compared to pre supplement (improvements in performance times, placebo = 37 s, 20 g = 158 s, 60 g = 134 s; 95% CI for differences, 47 s). CONCLUSION: Oral bovine colostrum supplementation at 20 g or 60 g/d provided a small but significant improvement in time trial performance in cyclists after a 2-h ride at 65% VO2max.

Administration, Oral↗

Use of immunoglobulin enriched bovine colostrum against oral challenge with enterohaemorrhagic Escherichia coli O157:H7 in mice.

An immunoglobulin enriched bovine colostrum preparation, IMMULAC (New Zealand Dairy Group, Cambridge, New Zealand), contains antibodies against various bacterial antigens. In the present study, we investigated the protective effects of a commercial bovine colostrum preparation against infections with enterohaemorrhagic Escherichia coli (EHEC) O157:H7 in a murine model. Balb/c mice were given drinking water containing streptomycin for 3 days before and following oral challenge with streptomycin-resistant EHEC O157:H7 strain (O157-SM(R)). In mice pretreated with streptomycin, EHEC O157:H7 maintained stable levels of bacterial colonization in the intestines for the 3-week experimental time period. Oral administration of colostrum resulted in rapid decrease in the bacteria numbers compared with administration of skim-milk. Colostrum showed no direct in vitro bactericidal properties against either EHEC O157:H7. When sections prepared from cecum walls of streptomycin-pretreated mice were incubated in vitro with EHEC O157:H7, the colostrum significantly prevented the attachment of the organisms to the sections when compared with skim-milk. These results indicate that oral administration of bovine colostrum effectively protects mice against food-borne infections by inhibiting bacterial attachment to the intestinal mucous membrane, colonization and growth in the intestinal tract.

Aging↗

Colostrum management in calves: effects of drenching vs. bottle feeding.

The objective was to examine whether the administration of colostrum by a drencher is a safe method to achieve satisfying immunoglobulin concentrations in newborn Holstein Friesian (HF)-calves. Twenty-one calves were fed 1 h postnatum with 2 l colostrum from their dam using a nipple bottle (group I); 15 calves were drenched with 4 l colostrum from their dam 1 h postnatum (group II). Thereafter, all calves were fed milk replacer exclusively. Serum concentration of total immunoglobulin [analysed by Sandwich-enzyme-linked immunosorbent assay (ELISA)] 24 h postnatum was higher in drenched calves (25.2 g/l; 12.8/52.2; median and 10/90 percentiles) compared with bottle-fed calves (14.1 g/l; 9.2/24.7). To characterize the kinetics of immunoglobulin absorption, 17 blood samples were taken frequently within 72 h postnatum using a jugular catheter in five calves fed 2 l colostrum by nipple bottle and five calves drenched with 4 l colostrum 1 h postnatum. A slightly delayed increase of serum immunoglobulin concentration (approximately 3 h) was obvious in drenched calves compared with bottle-fed calves, but drenched calves reached significantly higher immunoglobulin concentrations compared with bottle-fed calves. It is concluded that the proper application of colostrum by a drencher is a useful method for adequate colostral supply in newborn calves. The failure of the oesophageal groove reflex in drenched calves caused no clinical consequences.

Animals↗

Immune components (IgA, IgM, IgG, immune cells) of colostrum of Bangladeshi mothers.

BACKGROUND: Colostrum is the first defense for neonates. It is rich in immune components including immunoglobulins and viable immune cells. In the present study, human colostrum collected from 105 postpartum mothers was analyzed for its IgA, IgM, IgG levels, and peripheral immune cells. METHODS: Enzyme-linked immunosorbent assay was used to analyze the serum immunoglobulin concentrations. Immune cells were estimated by counting 200 cells. RESULTS: IgA was the dominant immunoglobulin and ranged from 2.84 to 8.69 g/L (mean 5.61 g/L). Mean IgM and IgG concentrations were 0.4 g/L (0.16-0.66 g/L) and 0.095 g/L (0.04-0.15 g/L), respectively. Neutrophil-macrophage (neu-mac) predominated in cell count (59%) followed by lymphocyte-plasma cells (lymph-plasma; 40%). The influence of maternal nutritional status, age, parity and income levels on the colostral immunological factors was studied. No significant association could be traced for immunoglobulin content, suggesting that maternal characteristics do not have any bearing on the immunoglobulin content of colostrum. Mean value of eosinophils was found to be higher among the underweight than the normal mothers (F= 3.143, r=-0.101). Maternal age was positively correlated with eosinophil (F= 3.296, r= 0.162). Concentration of neu-mac had a positive significant correlation with parity (t=-2.07, r= 0.205), while it was negatively significant for lymphocyte-plasma cells (t= 2.073, r=-0.101). However, the correlation coefficients of the immunologic parameters with other maternal characteristics were statistically insignificant. CONCLUSION: Colostrum has enough humoral and cellular elements to protect babies. Therefore, immune protection derived from breastfeeding depends on the immunoglobulin level of the colostrum as well as the amount of colostrum ingested.

Adolescent↗

Human colostrum contains IgA antibodies reactive to colonization factors I and II of enterotoxigenic Escherichia coli.

Diarrhea is an important cause of morbidity and mortality amongst infants of low socio-economic levels in developing countries and in travelers who visit such areas. Enterotoxigenic E. coli strains express two sets of virulence-associated factors: enterotoxins (heat-stable toxins or heat-labile toxins) and colonization factors. Studies have shown that breast-feeding protects infants against infectious diseases, such as diarrhea, as it presents a great variety of immunological components. The aim of this study was to analyze the reactivity of immunoglobulin A from human colostrum to colonization factor antigens I and II. The colostrum ability in preventing enterotoxigenic E. coli adhesion to Caco-2 cells was also evaluated. Colostrum samples were collected from 32 healthy women, and a human colostrum pool was prepared. Enterotoxigenic E. coli strains expressing colonization factor antigens I and II were utilized. The colostrum pool and individual samples showed variable antienterotoxigenic E. coli immunoglobulin A titers, that were reactive with colonization factor antigen I and CS1/CS3 (colonization factor antigen II). The human colostrum pool and individual samples inhibited enterotoxigenic E. coli colonization factor antigen I and II adhesion to Caco-2 cells, at variable levels, and this ability was a result of immunoglobulin A antibodies reactive to these colonization factors. The immunoglobulin A-depleted pool lost this inhibitory ability. As bacterial adhesion is the initial mechanism of enterotoxigenic E. coli infection, breast-feeding could protect the offspring against diarrhea caused by this agent.

Adolescent↗

Potential prophylactic value of bovine colostrum in necrotizing enterocolitis in neonates: an in vitro study on bacterial attachment, antibody levels and cytokine production.

Necrotizing enterocolitis (NEC) is an important disease of low birth-weight neonates. The immaturity of the gut mucosa may result in close contact between the host epithelium and microorganisms which are normally confined to the gut lumen. Damage of the mucosa due to endotoxin, cytokine production or other factors is believed to then occur. The aim of this study was to determine whether spray-dried bovine colostrum demonstrated potential in vitro as a prophylactic for NEC. Antiadherence was measured using a tissue culture assay and antibody levels against Enterobacteriaceae were determined by ELISA. The effect of bovine colostrum on the production of cytokines implicated in NEC was determined by a multiplex bead assay. Enterobacter cloacae, Klebsiella oxytoca, Escherichia coli, Serratia marcescens and Klebsiella pneumoniae ssp. pneumoniae were common in both NEC positive and NEC negative infants and IgA and IgG1 antibodies to these species were present in the bovine colostrum. Pretreatment with bovine colostrum produced a significant decrease (P<0.001) in attachment of bacteria to HT-29 cells. Bovine colostrum significantly increased the production of IL-8 in HT-29 cells and IL-8, IL-6 and TNF-alpha in THP-1 cells (P<0.001). The potential of bovine colostrum to increase the production of inflammatory mediators could limit its usefulness.

Animals↗

Identification of the complement regulatory protein CD59 in human colostrum and milk.

PROBLEM: Complement lytic activity has been demonstrated, and a potential for its activation is present in human colostrum and milk. This necessitates the presence of regulatory mechanisms protecting epithelial cells in the oropharynx and the gastrointestinal tract of the infant, the milk cellular elements, and bacteria colonizing the oropharynx and the gastrointestinal tract. Lactoferrin and C1 inhibitor have been attributed such a role. However, it is likely that additional protection against the cytolytic activity of the membrane attack complex is required. This has lead us to investigate the presence of the complement regulatory protein CD59 in human colostrum and milk, and to further characterize the source of secretion. METHOD: Samples of human colostrum and milk were obtained from volunteers at different stages of lactation, and separated into fat, skim milk, and milk cellular elements by centrifugation. Normal human mammary gland tissues were obtained from patients undergoing biopsy for benign conditions. SDS-PAGE and Western blotting, and an immuno dot-blot assay were used to identify CD59 in human milk. Immunohistochemistry was performed on all tissue samples and cytospins of the milk cellular elements, using monoclonal antibodies to CD59. RESULTS: CD59 was present in cell-free colostrum and milk as a 19-25 kDa glycoprotein. No variation in CD59 levels was detected between colostrum and milk. CD59 was present in great amounts in the cytoplasm and was highly expressed on the surface membrane on mammary gland acinar and ductal epithelial cells, while the milk cellular elements contained CD59 mainly in their cytoplasm. CONCLUSION: The complement regulatory protein CD59 present in cell-free human colostrum and milk may exert its effects both in the mammary gland and in the oropharynx and gastrointestinal tract of the infant. The lobuloalveolar epithelial cells in the mammary gland are the likely source of secretion.

Breast↗

Vitamin B12 in human colostrum and milk. Quantitation of the vitamin and its binder and the uptake of bound vitamin B12 by intestinal bacteria.

The concentration of vitamin B12 was measured by microbiological assay in 229 samples of normal human colostrum and milk taken at various stages of lactation. Colostrum obtained within 48 hours of delivery contains high concentrations of vitamin B12 (mean 2 431 pg/ml), but within a few days the levels fall to a range similar to the levels in normal serum. The vitamin B12 binding capacity of 111 samples of colostrum and milk was estimated by gel filtration or charcoal binding. Colostrum samples have a mean binding capacity of 72 ng/ml, while the binding capacity in milk is only one third of this value. The ability of a range of intestinal bacteria to take up colostrum-bound vitamin B12, was assessed. All the organisms took up free vitamin B12, but when the vitamin was bound in colostrum, there was little or no uptake even after 24 hours incubation.

Bacteria↗

Bile salt-stimulated lipase activity in human colostrum from mothers of infants of different gestational age and birthweight.

The bile salt-stimulated lipase (BSSL) activity was measured in milk collected at 3-4 days postpartum (colostrum) from 36 mothers divided into three groups according to gestational age and birthweight of their infants. BSSL activity changed with the length of gestation. Preterm colostrum presented a mean activity significantly higher than the term groups (small-for-gestational age and appropriate-for-gestational age) which had similar values. The ratio of BSSL activity to the estimated fat content was 6.33 in colostrum of mothers who delivered preterm and 4.20 in colostrum of both groups of term mothers. These data suggest that preterm colostrum has a higher fat digesting potential than term colostrum and that it is the gestational age rather than the adequacy or non-adequacy of birthweight to gestational age that may influence the BSSL activity.

Adult↗

IgG4 in human colostrum and human milk: continued local production or selective transport from serum.

Colostrum, mature milk, and paired plasma samples were obtained from 10 postpartum women who had not been previously studied. The geometric mean concentration of IgG4 in colostrum (3.3 micrograms/ml) was similar to the mean concentration in mature milk (3.0 micrograms/ml). The arithmetic mean for the percent of IgG = IgG4 was 10.3 +/- 3.3% for colostrum, 10.3 +/- 3.1% for mature milk, 2.6 +/- 0.3% for early plasma, and 1.7 +/- 0.3% for later plasma. Local mammary production of immunoglobulin was determined by subtracting the estimated serum contribution from the measured concentration in colostrum or milk. Evidence for local mammary production of IgG4 was found in 5 of 10 colostrum samples and 8 of 10 mature milk samples. These observations indicate that the previously observed selective enrichment of IgG4 in colostrum is also true for mature milk. These are the first studies suggesting continued local production of any immunoglobulin other than IgA in mature human breast milk.

Colostrum↗