Lymphocytes In Gut-associated Lymphoid Tissue Research Articles

Effects of parenteral nutrition supplemented with beta‐hydroxy‐beta‐methylbutyrate on gut‐associated lymphoid tissue and morphology in mice

AbstractBackgroundParenteral nutrition (PN) without enteral nutrition (EN) leads to marked atrophy of gut‐associated lymphoid tissue (GALT), causing mucosal defense failure in both the gut and the extraintestinal mucosal system. We evaluated the effects of beta‐hydroxy‐beta‐methylbutyrate (HMB) on GALT and gut morphology in PN‐fed mice.MethodsExperiment 1: male Institute of Cancer Research mice were assigned to the Chow (n = 12), Control (standard PN: n = 10), or H600 and H2000 (PN containing 600 mg/kg or H2000 mg/kg body weight of Ca‐HMB: n = 12 and 10, respectively) groups. After 5 days of dietary manipulation, all mice were killed and the whole small intestine was harvested. GALT lymphocyte cell numbers and phenotypes of Peyer patch (PP), intraepithelial space, and lamina propria lymphocytes were evaluated. Experiment 2: 47 mice (Chow: n = 12; Control: n = 14; H600: n = 11; and H2000: n = 10) were fed for 5 days as in experiment 1. Proliferation and apoptosis of gut immune cells and mucosa, and protein expressions (mammalian target of rapamycin [mTOR], caspase‐3, and Bcl2) were evaluated in the small intestine.ResultsCompared with the Controls, the Chow and HMB groups showed significantly higher PP cell numbers, prevented gut mucosal atrophy, inhibited apoptosis of gut cells, and increased their proliferation in association with increased mTOR activity and Bcl2 expression.ConclusionHMB‐supplemented PN is a potentially novel method of preserving GALT mass and gut morphology in the absence of EN.

Effects of short-term fasting on gut-associated lymphoid tissue and intestinal morphology in mice

Summary Background & aims Modern management guidelines for surgical patients recommend that the fasting period before surgery be as short as possible. This study aimed to examine the influences of 24-h fasting, with and without water, on intestinal morphology, mucosal IgA levels and gut-associated lymphoid tissue (GALT) cells as a center of systemic mucosal immunity in mice. Methods Male Institute of Cancer Research mice (6-week-old, n = 24) were divided into three groups: ad libitum chow and water (Control), 24-h fasting without water (Fasted − water), and 24-h fasting with ad libitum water (Fasted + water). The Fasted + water group was used to examine possible influences of dehydration on the parameters measured. After a 24-h fasting, nasal and bronchoalveolar lavage were done under anesthesia. Then, the mice were killed by cardiac puncture. The whole small intestine was harvested and intestinal washing was obtained by lavage. Jejunal and ileal morphologies were evaluated by HE however, significant decreases were observed in IEL and LPL numbers in the Fasted − water and Fasted + water groups as compared to the Control. Conclusions Regardless of water supply, 24 h of fasting causes gut atrophy, with marked GALT lymphocyte losses in the IE space and LP.

Expression of XBP1 in lymphocytes of the small intestine in rats under chronic social stress and modulation of intestinal microflora composition

The present study was conducted to investigate of the influence of chronic social stress and modulation of the composition of intestinal microflora on the distribution of Xbp(1+)-lymphocytes in the gut-associated lymphoid tissue of ileum of the rats. Structure of population of Xbp(1+)-cells has been studied by the analysis of serial histological sections using the method of indirect immunofluorescence with monoclonal antibodies to Xbp1 of rat. Chronic social stress development is accompanied with the reduction of total number of Xbp(1+)-lymphocytes in lymphoid structures of ileum (31% -3 fold reduction, p < 0.05), mostly expressed in lymphoidfollicles, and changes the concentration of Xbp1 protein in immunopositive cells. Modulation of the composition of intestinal microflora by antibiotics and probiotics under chronic social stress results in the increase of total number of Xbp1+ lymphocytes in gut-associated lymphoid tissue, the degree of it depends on the kind ofstress. The discovered alterations of Xbp1 expression under stress may be one of the triggers for development of autoimmune and inflammatory bowel diseases. Thus, increased understanding of the molecular actions and transcriptional networks regulated by XBP1 in immune cells may aid in the development of potential therapeutics targeting immune disorders.

Effects of a new immune-modulating diet enriched with whey-hydrolyzed peptide, fermented milk, and isomaltulose on gut associated lymphoid tissue in mice

Summary Aim To examine whether an enteral immune-modulating diet (IMD) enriched with whey-hydrolyzed peptide, fermented milk, and isomaltulose enhances gut associated lymphoid tissue (GALT) mass and function as compared with a standard enteral diet (STD). Methods Male ICR mice were randomized into the IMD ( n = 10) or STD ( n = 10) group. After 7 days of each feeding protocol, whole small intestines were harvested. GALT lymphocytes from Peyer's patches (PPs), intraepithelial spaces (IE) and the lamina propria (LP) were isolated, counted and subjected to phenotypic determination by flowcytometry (αβTCR+, γδTCR+, CD4+, CD8+ and B cells). In another set of mice (IMD; n = 13, STD; n = 13), immunoglobulin A (IgA) levels of small intestinal, nasal and broncho-alveolar washings were measured with ELISA. Results Total lymphocyte numbers in PPs and LP and absolute numbers of B cells in PPs, γδTCR + cells in IE and αβTCR+, γδTCR+, CD4+, and CD8+ cells in LP were significantly higher in the IMD than in the STD group. IgA levels of small intestinal washings were significantly higher in the IMD than in the STD group with no differences in respiratory tract IgA levels. Conclusion The new IMD increases GALT mass and gut IgA levels as compared with STD.

Influence of Adding Pyrroloquinoline Quinone to Parenteral Nutrition on Gut‐Associated Lymphoid Tissue

Experimental intravenous (IV) parenteral nutrition (PN) diminishes gut-associated lymphoid tissue (GALT) cell number and function. PN solution cannot maintain GALT at the same level as a normal diet, even when delivered intragastrically (IG). Previous studies demonstrated pyrroloquinoline quinone (PQQ)-deficient mice to be less immunologically responsive. Because standard (STD) PN solution lacks PQQ, PQQ supplementation may prevent PN-induced GALT changes. This study was designed to determine the influence of adding PQQ to PN on GALT. In experiment 1, mice (n = 32) were randomized to chow, IV-STD-PN, and IV-PQQ-PN groups. The chow group was fed chow with the same caloric content as PN. The IV-STD-PN group received STD-PN solution, whereas the IV-PQQ-PN group was given PQQ (3 mcg/d)-enriched PN by the IV route. After 5 days of feeding, lymphocytes were isolated from the Peyer's patch (PPs), intraepithelial space (IE), and lamina propria (LP) of the small intestine. GALT lymphocyte number and phenotype (αβTCR+, γδTCR+, CD4+, CD8+, B220+ cells) and intestinal immunoglobulin A (IgA) level were determined. In experiment 2, mice (n = 28) were randomized to IG-STD-PN or IG-PQQ-PN group. After IG nutrition supports, GALT mass and function were determined as in experiment 1. The IV-PQQ-PN group showed increased PP lymphocyte number and PP CD8+ cell number compared with the IV-STD PN group. The IG-PQQ-PN group had significantly greater PP lymphocyte number and PP CD4+ cell numbers than the IG-STD-PN group. Neither IV nor IG PQQ treatment raised IgA level. PQQ added to PN partly restores GALT mass, although its effects on GALT function remain unclear.

Effects of Adding Butyric Acid to PN on Gut‐Associated Lymphoid Tissue and Mucosal Immunoglobulin A Levels

Parenteral nutrition (PN) causes intestinal mucosal atrophy, gut-associated lymphoid tissue (GALT) atrophy and dysfunction, leading to impaired mucosal immunity and increased susceptibility to infectious complications. Therefore, new PN formulations are needed to maintain mucosal immunity. Short-chain fatty acids have been demonstrated to exert beneficial effects on the intestinal mucosa. We examined the effects of adding butyric acid to PN on GALT lymphocyte numbers, phenotypes, mucosal immunoglobulin A (IgA) levels, and intestinal morphology in mice. Male Institute of Cancer Research mice (n = 103) were randomized to receive either standard PN (S-PN), butyric acid-supplemented PN (Bu-PN), or ad libitum chow (control) groups. The mice were fed these respective diets for 5 days. In experiment 1, cells were isolated from Peyer's patches (PPs) to determine lymphocyte numbers and phenotypes (αβTCR(+), γδTCR(+), CD4(+), CD8(+), B220(+) cells). IgA levels in small intestinal washings were also measured. In experiment 2, IgA levels in respiratory tract (bronchoalveolar and nasal) washings were measured. In experiment 3, small intestinal morphology was evaluated. Lymphocyte yields from PPs and small intestinal, bronchoalveolar, and nasal washing IgA levels were all significantly lower in the S-PN group than in the control group. Bu-PN moderately, but significantly, restored PP lymphocyte numbers, as well as intestinal and bronchoalveolar IgA levels, as compared with S-PN. Villous height and crypt depth in the small intestine were significantly decreased in the S-PN group vs the control group, however Bu-PN restored intestinal morphology. A new PN formula containing butyric acid is feasible and would ameliorate PN-induced impairment of mucosal immunity.

Neutrophil Elastase Inhibitor Restores Gut Ischemia Reperfusion-Induced Impairment of Gut Immunity with Reduced Plasma Interleukin-6 Concentrations in Mice

Gut-associated lymphoid tissue (GALT) is regarded as central mucosa-associated lymphoid tissue that influences systemic mucosal immunity. Our previous study revealed that gut ischemia and reperfusion (I/R) reduces GALT lymphocyte numbers. Because gut hypoperfusion frequently occurs in trauma, shock, and surgery patients, establishment of a therapeutic method to preserve GALT mass after gut I/R may be important for the prevention of infections. We examined the effects of sivelestat sodium hydrate, a selective inhibitor of neutrophil elastase, on GALT mass and plasma cytokine concentrations in a murine gut I/R model. Seventy male ICR mice were randomized to the control (n = 34) or the sivelestat (n = 36) group. After intravenous cannulation of the animals, the superior mesenteric artery was occluded for 60 min. After reperfusion, physiologic saline or sivelestat 5 mg/kg hourly was infused for 24 h. Sixteen mice in the control and 22 in the sivelestat group were alive at 24 h. Twenty-six mice (n = 13 in each group) were chosen randomly for harvest of the small intestine. Lymphocytes from Peyer patches (PP), the intraepithelial space (IE), and the lamina propria (LP) were counted; and their phenotypes (αβT-cell receptor (TCR), γδTCR, CD4, CD8, B cell) were determined by flow cytometry. Cytokine concentrations (interleukin [IL]-6, IL-1β, IL-10) in the plasma and bronchoalveolar lavage fluid were measured by enzyme-linked immunosorbent assay. Sivelestat treatment did not improve survival but increased PP and IE lymphocyte numbers significantly and reduced the LP CD8(+) cell percentage and plasma IL-6 concentration compared with controls. There were no significant differences between the two groups in other cell phenotypes or cytokine concentrations. Sivelestat treatment after gut I/R may be useful for maintaining gut immunity and preventing systemic inflammatory responses.

Reversal of Parenteral Nutrition-induced Gut Mucosal Immunity Impairment With Small Amounts of a Complex Enteral Diet

Although parenteral nutrition (PN) prevents progressive malnutrition, lack of enteral nutrition (EN) during PN leads to gut associated lymphoid tissue (GALT) atrophy and dysfunction. Administering a small amount of EN with PN reportedly prevents increases in intestinal permeability. However, its effects on GALT remain unclear. We analyzed the minimum amount of EN required to preserve gut immunity during PN. Male Institute of Cancer Research mice underwent jugular vein catheter insertion and tube gastrostomy. They were randomized into four groups to receive isocaloric and isonitrogenous nutritional support with variable EN to PN ratios (EN 0, EN 33, EN 66, and EN 100). EN was provided with a complex enteral diet. After 5 days of feeding, the mice were killed and whole small intestines were harvested. GALT lymphocytes were isolated and counted. Their phenotypes were analyzed by flow cytometry. IgA levels of small intestinal washings were analyzed with enzyme-linked immunosorbent assay. Body weight changes did not differ between any two of the groups. Peyer's patch lymphocyte numbers increased in proportion to EN amount, whereas lamina propria lymphocyte numbers were significantly higher in the EN 100 than in the EN 0 group, with no marked increases in the EN 33 and EN 66 groups. Small intestinal IgA levels increased EN amount-dependently and reached a plateau at EN 66. A small amount of EN partially reverses PN-induced GALT changes, suggesting beneficial but limited effects on gut mucosal immunity.

Fish oil infusion reverses 5-fluorouracil-induced impairments in mucosal immunity in mice

Anticancer drugs frequently have deleterious effects on host defense against infection, limiting their clinical application. We previously demonstrated continuous infusion of 5-fluorouracil (5FU) to reduce gut associated lymphoid tissue (GALT) mass and secretory IgA levels. This study was designed to examine the effects of concomitant infusion of fish oil on gut mucosal immunity in mice receiving 5FU. Male ICR mice were randomized to the control (n=12), 5FU (n=12), or 5FU+FO (n=10) group. The 5FU and 5FU+FO groups received continuous IV infusion of 5FU at 10 mg/kg for 5 days. The 5FU+FO group was given a simultaneous infusion of 10 ml/kg of a 10% fish oil emulsion. The controls received normal saline at 0.3 ml/h. During these treatments, all mice were allowed free access to chow and water ad libitum. Then, the mice were sacrificed and GALT lymphocytes were isolated from Peyer's patches (PPs), the intraepithelial space (IE), and the lamina propria (LP). Small intestinal, nasal and broncho-alveolar (BALF) washings were also obtained. Lymphocyte yields from each site and phenotypes (CD4, CD8, alphabetaTCR, gammadeltaTCR, B220) were determined. IgA levels in the washings were measured with ELISA. The 5FU group had significantly lower IE and LP lymphocyte numbers and small intestinal and BALF IgA levels than the control group, with no differences in the percentages of any phenotypes. However, fish oil infusion restored IE and LP lymphocyte numbers and BALF IgA to control group levels. Fish oil infusion along with 5FU preserves GALT lymphocyte numbers and respiratory IgA levels.

Influence of Adding Fish Oil to Parenteral Nutrition on Gut‐Associated Lymphoid Tissue

Lack of enteral nutrition reduces gut-associated lymphoid tissue (GALT) mass and function, a mechanism underlying the increased morbidity of infectious complications in severely injured or critically ill patients. Strategies to restore parenteral nutrition (PN)-induced changes of GALT mass and function have been pursued. However, the influences of adding fish oil to PN on gut immunity remain to be clarified. Male Institute of Cancer Research (ICR) mice (n = 50) were randomized to 4 groups: ad libitum chow (chow), fat free PN (fat (-)-PN), PN + fish oil (FO-PN), and PN + safflower oil (SO-PN). The PN groups were given isocaloric and isonitrogenous PN solutions. The FO- and SO-PN groups received 20% of total calories from fat emulsions. After 5 days of feeding, lymphocytes from Peyer's patches (PPs), the intraepithelial space (IE), and the lamina propria (LP) of the entire small intestine were isolated. GALT lymphocyte numbers and phenotypes (CD4+, CD8+, alphabetaTCR+, gammadeltaTCR+, B220+ cells) were determined. Immunoglobulin A (IgA) levels of small intestinal washings were also measured by enzyme-linked immunosorbent assay. Another set of mice (n = 24) was used to determine plasma fatty acid compositions after feeding. Lymphocyte numbers from PPs and the LP and intestinal IgA levels were significantly lower in the PN groups than in the chow group, with no significant differences between any 2 PN groups. The FO- and SO-PN groups showed moderate recovery of IE cell numbers compared with the fat (-)-PN group. Omega-3 and omega-6 fatty acid levels were increased with fish and safflower oil additions, respectively, compared with the fat (-)-PN group. Adding fish oil to PN does not exacerbate PN-induced GALT changes but rather partially reverses these changes, with increased plasma omega-3 fatty acid levels.

LACK OF ENTERAL NUTRITION BLUNTS EXTRACELLULAR-REGULATED KINASE PHOSPHORYLATION IN GUT-ASSOCIATED LYMPHOID TISSUE

The mitogen-activated protein kinase (MAPK) family (extracellular-regulated kinase [ERK], p38, etc.) of signal transduction proteins includes important intracellular mediators of inflammation, playing critical roles in host defense. Phosphorylations of ERK and p38 are responsible for cell proliferation, cell differentiation, and cell death. We hypothesized that impaired gut-associated lymphoid tissue (GALT) function in the absence of enteral nutrition is associated with reduced MAPK phosphorylation in GALT cells. Fifty-three male Institute of Cancer Research mice were randomized into 3 groups; ad libitum chow, intragastric (i.g.)-TPN, and intravenous (i.v.)-TPN. TPN groups were administered a standard TPN solution. After 5 days of feeding, lymphocytes from Peyer patches (PPs), the lamina propria (LP) cells, and intraepithelial (IE) spaces in the small intestine were isolated. GALT lymphocyte numbers were determined. The lymphocytes were incubated with or without 50 ng/mL of phorbol myristate acetate (PMA) for 15 min, and phosphorylated ERK (p-ERK) and p38 (p-p38) levels were determined using laser scanning cytometry. In PP (GALT inductive site) lymphocytes, p-ERK was increased after PMA in all three groups. However, ERK phosphorylation in GALT effector sites (IE and LP) was enhanced only in the enteral groups. p38 phosphorylation was not increased in any GALT sites, in any of the three groups, in response to PMA. In another set of mice (n = 33), in vitro LP lymphocyte proliferation was assessed with BrdU with or without PMA. Cell proliferation was increased or maintained at high level with PMA in the i.g.-TPN and chow group, but remained low in the i.v.-TPN group. In conclusion, lack of enteral feeding blunts ERK activation and cell proliferation in response to PMA stimulation in GALT effector sites, which may be an important mechanism underlying reduced GALT function. The influence of nutrition on GALT p38 phosphorylation must be assessed with other types and dosages of stimulants.

Albumin Infusion After Reperfusion Prevents Gut Ischemia‐Reperfusion‐Induced Gut‐Associated Lymphoid Tissue Atrophy

Our recent study clarified that gut ischemia-reperfusion (I/R) causes gut-associated lymphoid tissue (GALT) mass atrophy, a possible mechanism for increased morbidity of infectious complications after severe surgical insults. Because albumin administration reportedly reduces hemorrhagic shock-induced lung injury, we hypothesized that albumin treatment prevents GALT atrophy due to gut I/R. Male mice (n = 37) were randomized to albumin, normal saline, and sham groups. All groups underwent jugular vein catheter insertion. The albumin and normal saline groups underwent 75-minute occlusion of the superior mesenteric artery. During gut ischemia, all mice received normal saline infusions at 1.0 mL/h. The albumin group was given 5% bovine serum albumin in normal saline at 1.0 mL/h for 60 minutes after reperfusion, whereas the normal saline group received 0.9% sodium chloride at 1.0 mL/h. The sham group underwent laparotomy only. Mice were killed on day 1 or 7, and the entire small intestine was harvested. GALT lymphocytes were isolated and counted. Their phenotypes (alphabetaTCR, gammadeltaTCR, CD4, CD8, B220) were determined by flow cytometry. On day 1, the gut I/R groups showed significantly lower total lymphocyte and B cell numbers in Peyer's patches and the lamina propria than the sham group. However, the albumin infusion partially but significantly restored these cell numbers. On day 7, there were no significant differences in any of the parameters measured among the 3 groups. Albumin infusion after a gut ischemic insult may maintain gut immunity by preventing GALT atrophy.

Effect of HIV-1 infection on lymphocyte proliferation in gut-associated lymphoid tissue.

To determine the change in the percentage of proliferative and activated lymphocytes in gut-associated lymphoid tissue (GALT) in HIV-1-infected subjects compared with that in uninfected controls. We measured the percentage of proliferative (Ki-67+) and activated (CD-69+, HLA-DR+, CD45RO+) lymphocytes from GALT and peripheral blood in chronically HIV-1-infected (12) and uninfected (9) individuals. The percentage of proliferative GALT CD4+ T cells was increased in HIV-1-infected control subjects compared with that in uninfected controls (p <.007). Based on immunohistochemical staining, proliferative T cells were principally located in the parafollicular area surrounding lymphoid aggregates. The percentage of activated GALT lymphocytes, however, was not significantly different in HIV-1-infected individuals, whereas it was significantly increased in the peripheral blood of HIV-1-infected individuals. The percentage of peripheral blood lymphocytes trafficking to the intestine was also not significantly different in HIV-1-infected individuals compared with that in uninfected controls. CD4+ T cell proliferation in GALT is increased in HIV-1 infection without a significant alteration in the percentage of peripheral blood T cells trafficking to the gastrointestinal mucosa.

Effect of HIV-1 Infection on Lymphocyte Proliferation in Gut-Associated Lymphoid Tissue

Objective: To determine the change in the percentage of proliferative and activated lymphocytes in gut-associated lymphoid tissue (GALT) in HIV-1-infected subjects compared with that in uninfected controls. Methods: We measured the percentage of proliferative (Ki-67+) and activated (CD-69+, HLA-DR+, CD45RO+) lymphocytes from GALT and peripheral blood in chronically HIV-1-infected (12) and uninfected (9) individuals. Results: The percentage of proliferative GALT CD4+ T cells was increased in HIV-1-infected control subjects compared with that in uninfected controls (p < .007). Based on immunohistochemical staining, proliferative T cells were principally located in the parafollicular area surrounding lymphoid aggregates. The percentage of activated GALT lymphocytes, however, was not significantly different in HIV-1-infected individuals, whereas it was significantly increased in the peripheral blood of HIV-1infected individuals. The percentage of peripheral blood lymphocytes trafficking to the intestine was also not significantly different in HIV-1-infected individuals compared with that in uninfected controls. Conclusions: CD4+ T cell proliferation in GALT is increased in HIV-1 infection without a significant alteration in the percentage of peripheral blood T cells trafficking to the gastrointestinal mucosa.

IL-4 Protects Adult C57BL/6 Mice from ProlongedCryptosporidium parvumInfection: Analysis of CD4+αβ+IFN-γ+ and CD4+αβ+IL-4+ Lymphocytes in Gut-Associated Lymphoid Tissue During Resolution of Infection

AbstractResistance of adult C57BL/6 mice to severe Cryptosporidium parvum infection is dependent on CD4+αβ+ TCR lymphocytes. In this study, we demonstrated that treatment with anti-IFN-γ mAb extended oocyst excretion 18 days longer, and anti-IL-4 mAb extended oocyst excretion at least 11 days longer than isotype control mAb treatment. Analysis of the specific activity of anti-IFN-γ mAb present in treated mouse sera suggested that IFN-γ may have a limited role in the resolution phase of infection. Changes were also documented in numbers of CD4+αβ+IFN-γ+ and CD4+αβ+IL-4+ lymphocytes in Peyer’s patches and intraepithelium of adult C57BL/6 mice during resolution of C. parvum infection. Resistance to initial severe infection was associated with CD4+αβ+IFN-γ+ lymphocytes, and eventual resolution of infection was associated with CD4+αβ+IL-4+ lymphocytes. Analysis of cytokine expression following in vitro stimulation with C. parvum Ags during resolution of infection demonstrated consistent increases in CD4+αβ+IL-4+ lymphocytes, but not CD4+αβ+IFN-γ+ lymphocytes. The relevance of CD4+αβ+IL-4+ lymphocytes in protection against C. parvum was then evaluated in C57BL/6 IL-4 gene knockout mice (IL-4−/−). Adult IL-4−/− mice excreted oocysts in feces approximately 23 days longer than IL-4+/+ mice. Further, anti-IFN-γ mAb treatment increased the severity and the duration of infection in IL-4−/− mice compared with those in IL-4+/+ mice. Together, the data demonstrated that IFN-γ was important in the control of severity of infection, and either IFN-γ or IL-4 accelerated termination of infection. However, neither IL-4 nor IFN-γ was required for the final clearance of infection from the intestinal tract of adult mice.

Recovery of gut-associated lymphoid tissue and upper respiratory tract immunity after parenteral nutrition.

The authors characterize the recovery of parenteral nutrition-induced changes in gut-associated lymphoid tissue (GALT) and upper respiratory tract immunity with enteral nutrition and provide further information defining the effects of enteral feeding on mucosal immunity. The small intestine plays a prominent role in development and maintenance of mucosal immunity, both intestinal and extraintestinal, primarily through immunoglobulin A (IgA)-mediated mechanisms. Prior research has shown that mice fed total parenteral nutrition (TPN) have reduced GALT T and B cells, the cells responsible for IgA production, as well as impaired upper respiratory tract immunity to viral challenge of previously immunized animals. The recovery of TPN-induced changes in GALT and upper respiratory tract immunity after enteral refeeding is studied. Male institute of Cancer Research mice received 5 days of TPN followed by 0 to 4 days of chow. Small intestinal GALT was characterized by flow cytometry. In a second experiment, animals were immunized intranasally with moused-adapted influenza virus. Three weeks later, one group received a 5-day course of TPN followed by enteral refeeding for 5 days. A second group received TPN alone. Both groups were challenged with intranasal virus and killed 40 hours postchallenge to determine viral shedding from the upper respiratory tract. Animals fed TPN only had significantly fewer GALT lymphocytes compared with those chow-fed control subjects. Peyer's patch counts increased after a single day of refeeding, returning to normal levels by 48 hours. Lamina propria counts remained significantly depressed after 24 hours of refeeding, but also returned to normal after 48 hours of refeeding. The T-cell and B-cell populations mimicked total cell patterns. Lamina propria CD4+/CD8+ ratio returned to normal only after 72 hours of refeeding. None of the 9 animals refed enterally for 5 days were positive for viral shedding, compared with 8 of 12 matched TPN-fed animals. Enteral refeeding after TPN is associated with rapid repletion of GALT cellularity, initially within Peyer's patches and subsequently within the lamina propria. Refeeding corrects the impairment of IgA-mediated upper respiratory tract antiviral immunity occurring with TPN administration. This work further enhances the authors' knowledge of the underlying immunologic differences influenced by routes of nutrition.

Alimentary lymphomas in the horse

A series of 9 cases of primary diffuse alimentary lymphoma of the equine small intestine is described. Clinically, the principal effects were attributable to malabsorption and disordered alimentary function and several cases had severe anaemia; in four this was of the haemolytic type. Hypoalbuminaemia and elevated γ globulin levels were often present. The neoplasms were confined mainly to the small intestine and mesenteric lymph nodes, sometimes with some involvement of other lymph nodes as well. The large bowel was affected in one horse, but none of the cases showed detectable invasion of parenchymatous organs. Evidence is presented that some of these lymphomas were of follicle centre cell origin and it is suggested that they had probably arisen from B lymphocytes in gut-associated lymphoid tissue.

Cytotoxic effector cell function in organized gut-associated lymphoid tissue (GALT)

Lymphocytes from the organized gut-associated lymphoid tissues (GALT) of adult guinea pigs were examined for surface markers characteristic of T and B lymphocytes and for their capacity to function as effector cells in mitogen-induced cellular cytotoxicity (MICC) and antibody-dependent cellular cytotoxicity (ADCC) reactions. GALT lymphocytes formed rosettes with rabbit erythrocytes, a T-cell marker, and underwent proliferative responses in vitro in the presence of phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM). GALT lymphocytes were cytotoxic in vitro for erythrocyte and DBA mastocytoma targets in the presence of PHA. A population of GALT lymphocytes bound aggregated γ-globulin; however, they functioned poorly in ADCC reactions. Thus, organized GALT in the guinea pig contains lymphocytes capable of functioning in T-cell-dependent MICC reactions but either lacks the effector cell population which mediates ADCC or contains an effector cell which functions poorly in ADCC.