Enhanced immunological memory responses to Listeria monocytogenes in rodents, as measured by delayed-type hypersensitivity (DTH), adoptive transfer of DTH, and protective immunity, following Lactobacillus casei Shirota ingestion.

We have investigated the effect of orally administered Lactobacillus casei Shirota (L. casei) on immunological memory, as measured by delayed-type hypersensitivity (DTH) and acquired cellular resistance (ACR). The studies were performed in animal models in which the animals were rendered immune by a primary Listeria monocytogenes infection. It was shown that orally administered viable L. casei, and not heat-killed L. casei, enhanced significantly the antigen-specific DTH at 24 and 48 h in Wistar rats, Brown Norway rats, and BALB/c mice in a time- and dose-dependent fashion. L. casei had to be administered at least 3 days prior to the DTH assay at a daily dose of 10(9) CFU in order to induce significant effects. Long-term administration of 10(9) CFU of viable L. casei resulted in enhanced ACR, as demonstrated by reduced L. monocytogenes counts in the spleen and liver and diminished serum alanine aminotransferase activity after reinfection. Enhancement of cell-mediated immunological immune responses by L. casei was further established in an adoptive transfer study. Naïve recipient BALB/c mice, which were infused with nonadherent, immunized spleen cells from L. casei-fed donor BALB/c mice, showed significantly enhanced DTH responses at 24 and 48 h compared to recipient mice which received spleen cells from control donor mice. In conclusion, orally administered L. casei enhanced cell-mediated immunological memory responses. The effects relied on lactobacillus dose and viability as well as timing of supplementation and, further, appeared to be independent of host species or genetic background.

Modulation of delayed-type hypersensitivity and acquired cellular resistance by orally administered viable indigenous lactobacilli in Listeria monocytogenes infected Wistar rats.

AIMS: Various probiotic lactobacilli have been reported to modulate immunity. In this study we investigate the effects of viable indigenous Lactobacillus strains Utr-1, Utr-2 and Utr-3, on T cell-mediated immunological memory responses. METHODS AND RESULTS: In Listeria monocytogenes infected rats it was demonstrated that short-term daily ingestion of Lactobacillus strain Utr-3 significantly decreased delayed-type hypersensitivity (DTH) expression, whereas long-term, daily oral administration of Lactobacillus strain Utr-3 and Lactobacillus strain Utr-2 significantly enhanced acquired cellular resistance (ACR) towards Listeria re-infection. CONCLUSIONS: Our findings demonstrate that certain indigenous Lactobacillus strains are capable of modulating T cell-mediated immunity. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results support the importance of indigenous microflora analysis in probiotic lactobacilli studies.

Antagonistic activity of Lactobacillus casei strain shirota against gastrointestinal Listeria monocytogenes infection in rats.

In the present study, the effect of ingested viable Lactobacillus casei Shirota strain YIT9029 on oral infection with the enteric pathogen Listeria monocytogenes in Wistar rats was investigated. Rats were orally infected with 10(9) viable L. monocytogenes. Starting 3 days before the infection, rats received a daily dosage of 10(9) viable L. casei. It was shown that supplementation of L. casei significantly reduced the numbers of L. monocytogenes in stomach, caecum, faeces, spleen and liver, 2 days after L. monocytogenes infection. The number of L. monocytogenes in the mesenteric lymph nodes was not affected by the ingestion of L. casei. In comparison with control animals, the levels of the liver-specific alanine aminotransferase were lower in L. casei-fed rats. Histological analysis of spleen and liver revealed no differences between the experimental and control animals. In a parallel study with orally L. monocytogenes infected rats, it was shown that L. casei was able to increase cellular immunity significantly as determined with the delayed-type hypersensitivity response against heat-killed L. monocytogenes. In conclusion, in the present study it was shown that orally administered L. casei is able to enhance host resistance against oral L. monocytogenes infection. In the gastrointestinal tract, as well as in the spleen and liver, L. monocytogenes numbers were reduced. Furthermore, it is concluded that the enhancement of this anti-Listeria activity might be, at least partly, due to increased cell-mediated immunity.

Comparison of faecal Lactobacillus populations in experimental animals from different breeding facilities and possible consequences for probiotic studies.

AIMS: The effect of probiotic lactobacilli is likely dependent on the indigenous Lactobacillus strains in the intestinal tract. Since a substantial number of probiotic studies is performed in rodents, we compared the Lactobacillus strains of different rat and mouse populations in three animal facilities. METHODS AND RESULTS: SDS-PAGE and 16S rDNA analysis of cultured faecal lactobacilli revealed that different Lactobacillus strains were detected in genetically similar Wistar rats bred at different locations. Further, within the same animal facility host genetics did not affect the types of the predominant lactobacilli strains. CONCLUSIONS: Our results show that the environmental background of laboratory animals rather than host genetics determines the indigenous Lactobacillus strains that are found. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings underline the importance of microflora analysis in probiotic studies.

Enhanced antigen-specific delayed-type hypersensitivity and immunoglobulin G2b responses after oral administration of viable Lactobacillus casei YIT9029 in Wistar and Brown Norway rats.

In this study, the effects of orally administered viable Lactobacillus casei Shirota strain YIT9029 on the immunity parameters of Wistar and Brown Norway rats were examined. For this purpose, we used the Trichinella spiralis host resistance model. Two weeks before and during T. spiralis infection, rats were fed 10(9) viable L. casei bacteria 5 days per week. The T. spiralis-specific delayed-type hypersensitivity (DTH) response was significantly enhanced in both Wistar and Brown Norway rats given L. casei. In both rat strains fed L. casei, serum T. spiralis-specific immunoglobulin G2b (IgG2b) concentrations were also significantly increased. In the model, no significant effects of L. casei on larval counts or inflammatory reactions in the tongue musculature, body weights, or lymphoid organ weights were observed. Serum specific antibody responses, other than IgG2b, were not changed by feeding of L. casei. In contrast to L. casei, it was shown that orally administered Bifidobacterium breve or Bifidobacterium bifidum had no influence on the measured infection and immunity indices in the rat infection model. Since the rat DTH response is considered to be a manifestation of Th1 cell-mediated immunity and the IgG2b isotype has been associated with Th1 activity, it was concluded that Th1 cells could play an active role in the immunomodulatory effects of orally administered L. casei. Furthermore, our data do not indicate that the effect of oral supplementation with L. casei is dependent on the genetic background of the host.

Presence of germline and full-length IgA RNA transcripts among peritoneal B-1 cells.

Next to conventional B cells (or B-2 cells), peritoneal B-1 cells have been shown to contribute significantly to the production of IgA-secreting plasma cells in the gut. Evidence for this was mainly based on studies comprising manipulated animals, including lethally X-irradiated and transgenic mice. To examine the ability of peritoneal B-1 cells from untreated mice to switch actively to IgA in vivo, we performed RT-PCR analysis on FACS-sorted peritoneal B-cell subsets from untreated BALB/c mice in order to examine the presence of germline C alpha mRNA and mature C alpha mRNA transcripts. Germline C alpha and mature C alpha transcripts were readily detectable in peritoneal B-1 cells (defined as IgMbright/IgDdull), but not, or very little, in peritoneal B-2 cells (defined as IgMdull/IgDbright). Moreover, by subdividing the B-1-cell population in CD5+ B-1a cells and CD5- B-1b cells, it was shown that in vivo expression of germline C alpha and mature C alpha transcripts was largely restricted to the B-1b-cell lineage. These results indicate that peritoneal B-1 cells indeed are capable to switch to IgA under normal physiological conditions and hereby further support the view that B-1 cells contribute significantly to the mucosal IgA response, albeit this function appears to be restricted to the B-1b-cell subset.

The BCL-6 proto-oncogene controls germinal-centre formation and Th2-type inflammation.

Structural alterations of the promoter region of the BCL-6 proto-oncogene represent the most frequent genetic alteration associated with non-Hodgkin lymphoma, a malignancy often deriving from germinal-centre B cells. The BCL-6 gene encodes a zinc-finger transcriptional repressor normally expressed in both B cells and CD4+ T cells within germinal centres, but its precise function is unknown. We show that mice deficient in BCL-6 displayed normal B-cell, T-cell and lymphoid-organ development but have a selective defect in T-cell-dependent antibody responses. This defect included a complete lack of affinity maturation and was due to the inability of follicular B cells to proliferate and form germinal centres. In addition, BCL-6-deficient mice developed an inflammatory response in multiple organs characterized by infiltrations of eosinophils and IgE-bearing B lymphocytes typical of a Th2-mediated hyperimmune response. Thus, BCL-6 functions as a transcriptional switch that controls germinal centre formation and may also modulate specific T-cell-mediated responses. Altered expression of BCL-6 in lymphoma represents a deregulation of the pathway normally leading to B cell proliferation and germinal centre formation.

B-1 cells and their reactivity with the murine intestinal microflora.

IgA secreting cells located in the lamina propria of the gut are a prominent feature of the mucosal immune system, which serves to protect the body from the continuous threat to infection by intestinal bacteria. In this review we summarize briefly the evidence that these IgA secreting cells have a dual origin and are derived either from conventional B cells or from B-1 cells. Furthermore, we show both at polyclonal and monoclonal levels that the major antigenic target of B-1 cell derived IgA are normal intestinal bacteria. Coating of intestinal bacteria with IgA is thought to result in immune exclusion, as shown for pathogenic bacteria. However, the bacterial microflora of the gut is an extremely stable ecosystem, despite the fact that the majority of intestinal bacteria are coated with IgA. We speculate here that these apparent contradictory functions of the humoral immune system, i.e. removal of bacteria and maintaining the normal gut flora might be exerted by IgA antibodies produced by the two B-cell lineages. The fixed and biased repertoire of B-1 cells might play a role in maintaining the normal intestinal flora. When pathogenic bacteria penetrate into the gut, conventional B cells may be induced in the Peyer's patches to produce high affinity, narrowly tuned IgA antibodies, leading to immune exclusion.

Motility disturbances in Graves' ophthalmopathy.

In a retrospective study of 30 patients with Graves' ophthalmopathy and 25 patients with exophthalmus due to other causes the patterns of ocular motility disturbances and the CT-findings were reviewed. In Graves' ophthalmopathy motility disturbances were found in 83% of the patients, in the control group 26% of the patients showed abnormal ocular motility. Enlarged eye muscles were found in 46% of the patients with Graves' ophthalmopathy and enlargement of the medial rectus muscle was the most consistent finding (42%). A clinical correlation was made between the severity of eye movement restriction and the appearance of the muscle on CT-scan.

Miniature optically immersed thermistor bolometer arrays.

Immersed thermistor bolometers have been in use since 1958 as sensors for ir horizon scanners employed in attitude control of earth-orbiting vehicles.(1) These detectors usually use a single small thermistor flake optically immersed in an antireflection coated pure germanium or silicon hemisphere or hyperhemisphere.(2) Studies of the earth's atmospheric horizon from orbiting vehicles indicate that the most useful radiant power lies in the carbon dioxide spectrum near 15 micro and in the rotational water bands of atmospheric moisture in the spectrum beyond 20 micro.(3) These atmospheric constituents produce high ir optical density and hence provide small angle horizon resolution. Carbon dioxide has the additional advantage, by being uniformly distributed in the atmosphere, of providing a stable horizon. The purpose of this paper is to describe briefly two types of five-element linear arrays of thermistor flakes optically immersed in germanium and silicon lenses (Fig. 1). These detecrs were designed for an advanced horizon definition study program at NASA-Langley Research Center.(4) Germanium immersion is employed for best detectivity in the carbon dioxide spectrum from 14 micro to 16 micro and silicon for the spectrum beyond 20 micro.