A variation in the formation of the median nerve: communicating branch between the musculocutaneous and median nerves in man.

We encountered variation in the formation of the median nerve in a 66-year-old male cadaver during dissection of the upper extremity of 20 adult cadavers. The dissections were made at the Department of Cellular Biology and Anatomy, Louisiana State University Medical Center. The median nerve was formed by fusion of four branches, three of them coming from the lateral cord and one from the medial cord. The normal radix from the lateral cord followed a very close oblique course over the axillary artery. The first unusual radix to the median nerve had an anastomoses from the musculocutaneous nerve to the median nerve in the proximal part of the left arm. The second unusual radix also came from the musculocutaneous nerve after it had pierced the coracobrachialis muscle and then joined with the median nerve. These kinds of variations are vulnerable to damage in radical neck dissection and other surgical operations of the axilla and upper arm. The communicating branch can be explained on the basis of its embryologic development and also ought to be distinguished from the other nerve variations in the upper extremity. The aim of this paper is to provide additional information for the classification of previously found communications between the musculocutaneous and median nerves.

The anastomotic artery connecting the axillary or brachial artery to one of the forearm arteries.

A vessel connecting the axillary or brachial artery to one of the forearm arteries was found in a 65 year old male cadaver, during the gross anatomy dissection of the upper extremity of 20 adult cadavers at the Department of Cellular Biology and Anatomy, Louisiana State University Medical Center. The right radial artery originated from the brachial artery nearly at the usual level and was connected to the axillary or brachial artery by a long slender anastomotic artery (vasa aberrantia). The anastomotic artery coursed under the medial side of the biceps muscle between the median and musculocutaneous nerves, and gave off two muscular branches to the biceps muscle. The anastomotic artery coursed between the median and musculocutaneous nerves in the arm, it passed to the forearm under the bicipital aponeurosis and connected the main radial artery on the radial side of the forearm. The anastomotic artery can be explained on the basis of its embryologic development and also ought to be distinguished from the other common arterial variations in the upper extremity.

Developmental aspects of intestinal intraepithelial and lamina propria lymphocytes in the rat following placental and lactational exposure to ethanol.

Fetal and lactational exposure to alcohol can induce impairments to the immune system and lead to decreased resistance to certain infectious agents. Morphometric procedures were used to quantify changes induced by maternal ethanol consumption in the gut-associated lymphoid tissue of rat pups. Rats were pair-fed with ethanol-containing or isocaloric control liquid diets formulated for pregnant and lactating animals from day 1 of pregnancy and throughout the lactation periods. Pups were weaned and placed on control liquid diet on post-natal day 21. Intraepithelial and lamina propria lymphocytes and macrophages were evaluated on post-natal days 14, 18, and 25. Lower thymus weights were observed in the ethanol-exposed pups on post-natal days 14 and 18 and lower total thymocyte counts on post-natal day 14. On post-natal day 14, T cells, T cytotoxic cells, IgA plasma cells, and macrophages were decreased in the ileal epithelial and lamina propria areas in the ethanol-exposed, compared to the pair-fed, pups. No differences for any of the above cell counts were found in the jejunum on post-natal day 14. On post-natal day 18, macrophages were still decreased in the ileum in the ethanol-treated pups, compared to pair-fed animals. No differences were found in T cells, T cytotoxic cells, and IgA lymphocytes between groups in either the jejunum or ileum on post-natal days 18 and 25. This study suggests that fetal and lactational exposure to ethanol has some effects on the development or influx of intraepithelial and lamina propria leukocytes and that the changes are most pronounced in early neonatal life.

Arterial, neural and muscular variations in the upper limbs of a single cadaver.

Arterial, neural and muscular variations were observed in both upper limbs of a female adult cadaver during routine student dissection. Two superficial radial arteries were observed, which originated from the brachial and axillary arteries in the left and right upper limbs respectively. In the right upper limb, an ulnar nerve with a lateral root from the lateral cord of the brachial plexus in the axilla and, in the left forearm, an accessory muscle belly inserting into the muscle belly of the flexor digitorum superficialis were also observed. Formation of these variations is discussed on the basis of the embryological development of the upper limb structures. The clinical importance of the variations is emphasized.

Second generation effects of maternal ethanol consumption on immunity to Trichinella spiralis in female rats.

The deleterious effects of maternal ethanol consumption on neonatal immune development and early immune responses has been well documented. However, the effects of such neonatal exposure to maternally consumed ethanol on the neonates' immune responses in their adult life, especially in combination with additional ethanol exposure, has received little attention. For these experiments, female rats were fed on either 6% ethanol or pair-fed isocaloric control Lieber-DeCarli liquid diets for 30 days prior to, and during, pregnancy and lactation. One day after weaning their pups, the mothers were infected with 1000 Trichinella spiralis larvae, and maintained on diets for an additional 20 days. At this time, they were challenged with 2000 T. spiralis larvae, killed 3 days later, and their immune status determined. These animals served as the first generation alcohol animals. Their female offspring served as the experimental second generation animals. These animals received maternal ethanol during pregnancy and lactation and control diet during their juvenile period (from weaning to 90 days of age). They were then subjected to a schedule of ethanol or pairfeeding, identical to the first generation dams. Two groups of second generation animals were established: Group 1 was exposed to ethanol during their dam's pregnancy and lactation periods only, with no subsequent ethanol treatment; Group 2 received ethanol during their dam's pregnancy and lactation periods and then again throughout their adult experimental period. Our previous studies showed only minimal changes following a secondary challenge in T. spiralis-immunized rats; however, neonates born to alcohol-consuming mothers did show some depressed secondary immune responses when challenged soon after weaning. We chose to use a secondary immune challenge to assess further immune alterations in second generation adult animals. No differences between any of the ethanol and pair-fed groups were observed in intestinal worm burdens, which is similar to data previously reported for adult alcohol-consuming animals. However, second generation group 2 animals demonstrated significantly reduced proliferation responses to T. spiralis antigen and Concanavalin A (Con A) stimulation relative to the ethanol first generation and to the second generation Group 1 animals. This group also demonstrated significantly lower absorbencies in the ELISA assay for specific IgM and IgG anti-T. spiralis antibodies than the pair-fed, ethanol first and second generation Group 1 animals. The proportion of total T cells and cytotoxic T cells was significantly lower and the proportion of natural killer cells was elevated in both second generation ethanol Groups 1 and 2 relative to the ethanol first generation and pair-fed groups. In addition, Group 2 second generation animals showed significantly lower proportions of total leukocytes and T cells than Group 1 second generation animals. Although secondary immune responses to T. spiralis infection were not altered in rats exposed to ethanol only as adults, exposure to maternal ethanol does affect some specific immune responses in second generation adult life and maternal exposure may exert cumulative immune effects in concert with later consumption of ethanol by offspring born to alcoholic mothers.

The temporal relationship among anti-parasite immune elements expressed during the early phase of infection of the rat with Trichinella spiralis.

Immunological parameters were measured during the first 20 days of infection with Trichinella spiralis in the rat. Expulsion of adult worms was complete by day 15 postinfection. Eosinophil and neutrophil numbers rose in the blood of infected rats above preinfection levels on days 3 and 6, respectively, and remained high to day 20 postinfection. Release of cytokines by Trichinella-antigen-stimulated mesenteric lymph node cells was measured, and a significant elevation in interferon (IFN)-gamma release was detected during the early stage of infection. Although initiated later, interleukin (IL)-10 release showed a pattern similar to IFN-gamma. Biphasic release of IL-5 was seen with significant elevation above the preinfection level on day 3 and after day 6 postinfection to the end of the study. IL-4 and IL-2 showed biphasic secretion as well, with the level of IL-4 high in the early and middle part of infection, while the level of IL-2 was detectable only at days 2, 3 and 6 postinfection. Serum anti-parasite IgE rose above preinfection levels after day 6 postinfection. Anti-parasite Ig-positive mesenteric lymph node (MLN) cells were evident by day 3 postinfection for IgM, and day 9 postinfection for IgA and total IgG. The number of Ig-positive MLN cells for all antibody classes returned to preinfection levels by day 20 postinfection. Evaluation of the temporal interactions of the key anti-parasite immune components with which the host engages Trichinella shows a complex interplay between Th1 and Th2 helper subsets.

Ethanol consumption suppresses cell-mediated inflammatory responses and increases T-helper type 2 cytokine secretion in Trichinella spiralis-infected rats.

Alcohol consumption has been shown to be associated with immune suppression and immune modulation. In this study, the effects of ethanol ingestion on the host immune responses to Trichinella spiralis infection and the subsequent secretion of T-helper cell-associated cytokines were investigated in rats. At the early phase of T. spiralis infection, ethanol-fed animals showed decreased numbers of blood neutrophils and eosinophils, and a decreased secretion of interferon-gamma (IFN-gamma) by mesenteric lymph node cells, compared with pair-fed controls. Suppression of this early inflammatory response to infection in the ethanol-treated groups resulted in a slower rate of expulsion of intestinal adult worms and a higher fecundity rate for female worms, compared with pair-fed controls. A dramatic decrease in blood neutrophils in the ethanol-treated groups was also manifested on day 9 postinfection. At that time, mesenteric lymph node cells from the ethanol-treated groups secreted higher amounts of mast cell proliferation-enhancing activity, which has been shown to contain T-helper type 2-associated cytokines. At the later phase of infection (day 12 to 20 day postinfection), ethanol-treated animals contained higher numbers of blood eosinophils and secreted an increased amount of interleukin-5 and mast cell proliferation-enhancing activity, compared with pair-fed controls. Although there was a slight rise with time after infection, the level of serum corticosterone was not significantly increased in the ethanol groups. Therefore, it is not likely that the observed immune modulations caused by ethanol consumption, especially in the early phase of infection, is the effect of elevated levels of corticosterone in the circulation. The present study found that ethanol consumption suppressed the initial amount of interferon-gamma secretion and inflammatory response, and may have directly or indirectly led to an enhancement of the secretion of T-helper type 2-type cytokines later in the primary immune response to T. spiralis infection.

Preliminary study of how alcohol consumption during pregnancy affects immune components in breast milk and blood of postpartum women.

Human milk has been shown to contain numerous immune components that can potentially protect the infant during the period before its own immune system is completely developed. Alcohol consumption in both experimental animals and humans has been associated with alterations to a number of immune parameters. We have investigated the possibility that alcohol consumption during pregnancy alters certain immune components in day 3 postpartum breast milk and peripheral blood of women. Our study group consisted of 10 alcoholic beverage drinkers (moderate to heavy, most of whom smoked a 1/2-1 pack of cigarettes per day), 15 non-drinking/non-smoking controls, and 10 non-drinking/smokers (1/2-1 pack per day) controls. The immune parameters measured in these otherwise healthy women were: (1) percentage and absolute number of the various subsets of leukocytes; (2) percentage of T cells, B cells, T helper and cytotoxic/suppressors subsets, and natural killer cells; (3) levels of the cytokines IL-6, IL-8 and TNF-alpha; (4) levels of IgA in milk and IgG in serum. Milk from the alcohol group contained an elevated amount of IL-8 as compared with milk from non-smoker controls; however, it did not differ statistically from that of the smoker controls. Blood from the alcohol group showed an increased level of IL-8 when compared with that from both smoker and non-smoker controls. The total number of leukocytes in milk was elevated in milk from the alcohol group as compared to both the smoker and non-smoker control groups. In the leukocyte component of milk, neutrophils predominate and are responsible for the elevation in total number of cells, as both lymphocyte and macrophage populations did not differ from those of the controls. For lymphocytes, B cells were also increased in blood of the smokers as compared with the alcohol and non-smokers controls. There were no statistical differences in any of the other immune parameters tested among the three groups. The present study found that alcohol consumption during pregnancy could modulate the production of IL-8 and infiltration of certain leukocytes in milk and blood of postpartum women. Some of these alterations were also evident in the smoker controls and thus could not be attributed to alcohol consumption alone.

Effects of maternal ethanol consumption on the subsequent development of immunity to Trichinella spiralis in rat neonates.

The immune response of rat pups to the intestinal parasite Trichinella spiralis was studied to determine if maternal pre- and/or postnatal ethanol consumption affected neonatal immune responses. Female rats were fed ethanol-containing (36% of calories) or pair-fed control liquid diets and include groups that were maintained on ethanol as follows: group 1, from day 1 of pregnancy through weaning and whose pups were then placed on ethanol to sacrifice; group 2, from day 1 of pregnancy through lactation; group 3, from day 1 of pregnancy through pup delivery; and group 4, from day 1 of lactation through weaning. A parallel group of animals was pair-fed isocaloric control diet until sacrifice. The pups of all litters were immunized orally with 500 L1 (T. spiralis) larva 5 days after weaning. To examine the effects of maternal ethanol on primary immune responses, one-fourth of the pups from each litter were sacrificed on days 10 and 20 after immunization. To examine the effects on neonatal secondary immune responses, the remaining pups were challenged with 1,000 larva 30 days after the initial immunization and sacrificed either 3 or 8 days after challenge. At the time of sacrifice, blood samples were collected, the intestine removed to determine T. spiralis worm burdens, and suspensions of mesenteric lymph node (MLN) cells prepared. Intestinal worm counts and serum levels of anti-T. spiralis IgM and IgG antibodies, interleukin-2 (IL-2), and tumor necrosis factor (TNF) were determined. In vitro proliferation responses of MLN cells to T. spiralis antigen and to the mitogen concanavalin A (Con A) were also examined. Pups from groups 1 to 3 demonstrated significantly higher intestinal worm counts (decreased immunity) than the pair-fed controls at the day 20 primary immune response sacrifice, and pups from group 1 had significantly higher worm counts at day 3 after a secondary immune challenge. Pups of dams from groups 1, 3, and 4 had significantly lower IgM antibody titers at the day 20 primary immune response sacrifice. All experimental ethanol groups (1 to 4) demonstrated significantly lower IgG antibody titers than that observed in pair-fed control pups at the 20-day sacrifice. IgM antibody titers showed significant reductions for ethanol-treated groups at 3 and 8 days after T. spiralis secondary challenge. In addition, IgG antibody titers were also significantly reduced for all alcohol groups at 3 and 8 days during the secondary immune response. Serum IL-2 and TNF levels were significantly lower in all experimental ethanol groups (1 to 4) relative to pair-fed controls at day 20 during a primary immune response, and IL-2 levels at 3 days postchallenge were lower in groups 2 to 4 after a secondary immune challenge. MLN proliferation responses to antigen and Con A were significantly reduced in ethanol groups 1 to 3 and to Con A in group 4 at day 10 after a primary immune challenge. Ethanol group 3 pups also demonstrated a reduced response to antigen at day 20. For animals undergoing a secondary immune response, ethanol group 2 demonstrated a reduced response to antigen at day 3, whereas groups 2 and 4 showed increased reactivity to antigen at days 3 and 8 postchallenge. These results show that maternal ethanol consumption diminishes the capacity of neonates to respond to T. spiralis antigen and that the depressed immune response involves T- and B-cell-mediated reactions and also affects the production of certain cytokines. These results also suggest that the diminished immune responses are increased with longer periods of maternal and neonatal exposure to ethanol.

Effect of maternal ethanol consumption on in vitro tumor necrosis factor, interleukin-6 and interleukin-2 production by rat milk and blood leukocytes.

It has been shown that the transfer of immunity via lactation plays an important role in providing early protection to the neonate. Maternal ethanol consumption also results in a reduced transfer of immunity to their neonates against a Trichinella spiralis infection. Because of the known presence of cytokines in milk and their important role in inflammation, we tested the effects of maternal ethanol consumption on cytokine production by milk and blood cells from T. spiralis-infected rats. With T. spiralis antigen, Concanavalin A (Con A) or lipopolysaccharide stimulation, milk cells from both ethanol-treated and pair-fed groups were capable of producing tumor necrosis factor (TNF), interleukin (IL)-6 and IL-2. There were no differences between groups for TNF or IL-6 production by milk cells. Milk cells from the ethanol group produced a significantly higher amount of IL-2 upon Con A stimulation, as compared with that from the pair-fed group (16 +/- 4 units/10(6) cells vs. 4 +/- 1 units/10(6) cells, p < 0.05). After stimulation with Con A, blood cells from the ethanol group produced significantly lower amounts of TNF (17 +/- 15 units/10(6) cells) than that from the pair-fed group (102 +/- 64 units/10(6) cells, p < 0.05). The amount of TNF and IL-6 produced by milk cells was significantly lower, as compared with that produced by blood cells. This study suggests that ethanol consumption has some modulatory effects on cytokine production by milk and blood cells.

In vitro TNF, IL-6 and IL-2 production by rat milk cells following Trichinella spiralis infection.

Trichinella spiralis-specific immunity can be transferred from immune mothers to suckling neonates via lactation, suggesting that milk from immune dams contains specific factors to T. spiralis. TNF and IL-6 are important cytokines in inflammatory processes, and IL-2 is essential in lymphocyte activation. Using cell line bioassays, we examined the capacity of rat milk mononuclear cells from immune and non-immune control dams to produce these cytokines following in vitro stimulation with mitogens or T. spiralis antigen. Milk cells were capable of producing IL-2, IL-6 and TNF upon Con A stimulation, and TNF and IL-6 upon LPS stimulation. The amount of these cytokines produced by mitogen-stimulated milk cells from T. spiralis-infected rats was similar to that produced by non-infected controls, although lower than that of corresponding blood mononuclear cells. Upon stimulation with T. spiralis antigen, milk cells from infected rats produced a significantly higher amount of TNF (158 +/- 39 U/10(6) cells) compared to non-infected controls (16 +/- 6 U/10(6) cells, P < 0.01), and also higher than that of the corresponding blood cells (60 +/- 10 U/10(6) cells, P < 0.01). Only small amounts of IL-6 and no IL-2 was secreted by milk cells from control or infected groups after stimulation with antigen. This study shows that rat milk cells are capable of synthesizing cytokines, and TNF produced by immune mothers may play a role in augmenting the neonate resistance to T. spiralis infection.

Effects of levamisole on ethanol-induced suppression of lactational immune transfer in rats.

Maternal ethanol consumption in rats has been shown to inhibit lactational transfer of immunity to Trichinella spiralis (T. spiralis) from dams to their neonates. The purpose of this study was to determine if this depressed immune transfer could be altered by treating the dams with a known immunostimulatory drug during pregnancy and lactation. Groups of female rats were fed ethanol-containing or were pair-fed isocaloric control liquid diets for 30 days, infected orally with 1,000 T. spiralis larva, and then continued on diet for 10 days to allow the adult worms to establish. The animals were placed on chow diets (maximum 5 days) and mated 1 to 1 with males. On day 1 of pregnancy the animals were returned to their respective liquid diets through pregnancy and lactation. One-half of the ethanol-treated animals was given 15 mg/kg body weight of levamisole in the diet beginning on day 10 of pregnancy and continuing until day 17 of lactation. On day 19 of lactation, pups from all experimental groups were challenged orally with 200 T. spiralis larva, and killed at 3 or 8 days postchallenge. Assays for intestinal worm burdens, IgG anti-T. spiralis serum antibodies, and mesenteric lymph node cell proliferation were conducted. At both sacrifice periods, pups from ethanol-treated animals showed significantly higher intestinal worm counts (decreased immunity) and significantly lower titers of specific antibodies than the pups of pair-fed animals or pups of animals receiving levamisole in addition to ethanol.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of maternal alcohol consumption on milk and blood lymphocytes and IgG antibody levels from Trichinella spiralis-infected rats.

Immunity to the intestinal parasite Trichinella spiralis can be transferred from the mother to the neonate during lactation. Previous studies in our laboratory showed that the passage of immunity to pups from ethanol-treated dams was depressed. This study examined the effect of ethanol consumption during pregnancy and lactation on the T. spiralis-associated immune components in milk and blood. Groups of female rats were fed either ethanol-containing or isocaloric liquid diets for 30 days before T. spiralis infection, mated and maintained on corresponding diets through pregnancy and lactation. Two-color flow cytometric analysis was performed for lymphocyte populations, enzyme-linked immunoabsorbent assay for specific IgG, and radial immunodiffusion assay for total IgG. The percentage of total T cells and their subsets, T helper cells and T cytotoxic/suppressor cells in milk and those in blood were similar between pair-fed and ethanol-treated animals. However, the percentage of natural killer cells in milk from ethanol animals was significantly reduced compared with the pair-fed group (33% vs. 54%). The percentage of activated or memory type T helper cell subset (OX2-W3/25+) was significantly increased in the blood of the ethanol-treated group. Pair-fed animals showed higher T. spiralis-specific IgG antibody levels in both in milk and blood compared with ethanol-treated animals. In ethanol-treated animals, specific IgG levels and total IgG concentration in milk were significantly lower than those in blood, whereas in pair-fed animals, only total IgG concentration in milk was lower than that in blood. This study indicates that ethanol consumption during pregnancy and lactation alters the maternal immune system.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of lymphocyte subsets in rat milk from normal and Trichinella spiralis-infected rats.

We have shown that T. spiralis-specific T lymphocytes can mediate maternal-to-neonatal immunity during lactation. This study addresses the change of lymphocyte populations in rat milk during normal and disease conditions. Two color flow cytometric analysis was performed for milk lymphocytes. T cells (OX19+) made up 45% of rat milk lymphocyte population. T helper cells (Th) composed 35% of total T cells while T cytotoxic/suppressor (Tcs) cells constituted 34%, giving a Th/Tcs ratio of 1.03. The corresponding ratio Th/Tcs in peripheral blood was 2.8. Approximately 21% of OX8+ cells in rat milk were OX19- natural killer (NK) cells. When using the monoclonal antibody 3.2.3 (NKR-P1), 43% of lymphocytes in control rat milk and 14% of blood lymphocytes were NK cells. This indicates a selective passage of these cells into milk. In T. spiralis-immunized rats, the percentage of total T cells was slightly decreased; however, Th and Tcs cells were consistent as compared to control milk. The percentage of NK cells (OX8+OX19-) in milk from T. spiralis-immunized rats was significantly higher than that from control milk (65% vs. 21%, respectively, P less than 0.01). This result was confirmed using the monoclonal antibody 3.2.3 which showed that milk from immunized rats contained 63% NK cells compared to 43% in normal milk (P less than 0.01). This study suggests that NK cells are selectively passaged into rat milk and T. spiralis infection induces an increase of NK cells in milk.

The effects of maternal ethanol consumption on lactational transfer of immunity to Trichinella spiralis in rats.

Transient immunity to the intestinal parasite Trichinella spiralis can be transferred from the mother to the neonate during lactation. The goal of this study was to determine whether maternal ingestion of ethanol during pregnancy and lactation inhibited expression of anti- T. spiralis immunity in nursing pups. Groups of female rats were infected with 1000 T. spiralis L1 larva, mated, and fed either ethanol-containing or isocaloric liquid diets and maintained on diets through pregnancy and lactation or were fed the liquid diets for 30 days before T. spiralis infection, mated, and maintained on diets through pregnancy and lactation. Pups were challenged orally with 200 T. spiralis larva at 14 days postdelivery (preweaning period) or 21 days postdelivery (postweaning period) and were sacrificed either 3 or 8 days after respective challenge. Intestinal worm counts and serum titers of anti-T. spiralis IgG antibodies were determined for each pup. No difference in the number of intestinal worms between pups of ethanol-treated and pair-fed dams that received ethanol diet after T. spiralis infection was observed in the preweaning period. This was also true of pups from the dams sacrificed at 3 days after challenge in the postweaning period. However, similar pups sacrificed at 8 days after challenge showed significantly higher worm counts (decreased immunity) relative to their pair-fed controls. Pups of dams that received ethanol containing diet 30 days prior to T. spiralis-infection showed significantly higher numbers of intestinal worms relative to pair-fed pups at both the preweaning and postweaning periods.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical comparison of T-cell and macrophage populations in mammary tissue of control and Trichinella spiralis-infected rats.

We have shown that antigen-specific T lymphocytes can mediate maternal-to-neonatal immunity during lactation. Present studies address the dynamics of lymphocyte accumulation in the mammary gland during normal and disease stimulated conditions. Monoclonal antibodies specific for total T cells, suppressor/cytotoxic and helper subsets, and macrophages were used in conjunction with immunohistochemistry to identify and count the individual cell types. In unstimulated mammary tissue, following a rise in T cells to maximal numbers in late pregnancy, the total number of T cells/high power field (HPF) was significantly diminished in early lactation and continued to decline to the late lactation period. Both the numbers of T cells/HPF located in the mammary alveolar epithelium and surrounding connective tissue were significantly reduced in early lactation as compared to late pregnancy. This indicates the possible passage of cells into the milk during lactation. Prior infection of the mother with Trichinella spiralis and a secondary challenge 48 h. before sacrifice caused a significant reduction in the number of T cells in the mammary tissue in early lactation as compared with unstimulated controls, indicating the possibility of an even greater outflow of T cells into milk. In controls, the T-suppressor/cytotoxic subtype showed a reduction in early lactation versus late pregnancy but showed no shifts in total cells/HPF during infection. The T-helper subtype in controls remained unchanged from late pregnancy to early lactation with a considerable decline in late lactation. However, the T-helper cells were significantly decreased in T. spiralis-treated animals as compared with noninfected controls in early lactation.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of maternal ethanol consumption on the distribution of leukocyte subsets in the lactating mammary gland of rats.

Significant immunological protection is provided to the newborn by the transfer of maternal leukocytes during nursing. The objective of this study was to determine if ethanol ingestion altered the distribution of T, B and accessory cells in the mammary glands of normal rats or in rats infected with Trichinella spiralis (T. spiralis). In addition, female rats were fed either Lieber/DeCarli regular (18% protein) or higher protein (25% protein) ethanol-containing liquid diets for pregnant or lactating rats or were pair-fed the corresponding isocaloric control diets to assess changes in cellular distribution due to dietary content. In the first experiment (short-term ethanol), animals were placed on either 18 or 25% protein diet (ethanol or pair-fed) on day 1 of pregnancy. In the second experiment (long-term ethanol), animals were placed on 18% protein diet (ethanol or pair-fed) for 30 days prior to mating, mated and then placed on either 25% protein diet (18%/25% animals) or kept on 18% protein diet (18%/18% animals) through pregnancy and lactation. In the third experiment (long-term ethanol, immunized), animals were fed 18%/25% protein diet (ethanol or pair-fed) as in experiment 2 and were infected intragastrically with 1000 T. spiralis larva on day 15 of pregnancy. All animals were sacrificed on day 2 of lactation, the mammary gland removed and weighed. Frozen sections of mammary gland from each animal were incubated with monoclonal antibodies directed against antigens associated with rat T cells (W3/13), suppressor/cytotoxic (OX8) or helper (W3/25) T cell subsets, macrophages (ED2) or rat IgA (B cells) and processed for peroxidase anti-peroxidase (PAP) immunocytochemistry. The total number of antigen positive leukocytes/high powered microscope field (hpf), intraepithelial leukocytes/hpf and alveolar connective tissue leukocytes/hpf was determined for each stain by counting 100 hpf and the data were compared by statistical analysis. Significant differences in mammary gland weight between ethanol-treated and pair-fed animals were observed in the long-term 18%/18% protein diet experiment in which animals fed ethanol had significantly lower mammary gland weights. Ethanol-treated short-term animals maintained on the 18% protein diet through pregnancy, showed decreased numbers of IgA+ B cells and W3/13+ T cells/hpf in the alveolar connective tissue compartment and this decrease was reflected in total cells/hpf. Long-term 18%/18% protein diet, ethanol-fed animal showed increases in total IgA+ B cells and W3/13+ T cells/hpf as compared with pair-fed controls.(ABSTRACT TRUNCATED AT 400 WORDS)

Role of T cell subsets in the maternal-to-neonatal transmission of immunity against Trichinella spiralis during lactation in rats.

We have previously demonstrated the maternal-to-neonatal transfer of immunity to T. spiralis during lactation and have shown that antigen-specific T lymphocytes, when injected into the mother or orally fed to neonates, can mediate this transfer. To further analyze the T cell subsets involved in conferring this protection, T lymphocytes were isolated from the mesenteric lymph nodes of syngeneic donor rats infected 4-6 days earlier with T. spiralis. The T cells were incubated in vitro with either mouse-anti-rat 0X8 or W3/25 monoclonal antibody, "panned" on plates coated with goat-anti-mouse Ig, and the non-adherent T helper or T cytotoxic/suppressor cells harvested. 100 x 10(6) T helper cells were injected i.v. into mothers once in early lactation and again two days prior to challenging their pups (200 T. spiralis larvae) at 2 weeks of age. This resulted in significant passage of immunity from the mothers to their suckling neonates, worm counts being 59% and 73% of control values 3 and 8 days post-challenge (P less than 0.01). Injection of T-cytotoxic/suppressor cells using the same regimen resulted in significant suppression of immunity in challenged pups, who retained worm counts that were 105% and 145% of control values at 3 and 8 days post-challenge. Synergy between recombined panned T-helper and T cytotoxic/suppressor cells without Ly1+2+3+ amplifier cells was tested by recombining non-adherent panned 0X8 and W3/25 cells. This resulted in no significant expressions of immunity in the pups when compared to controls. The presence of transferred maternal T cells within the neonate was evidenced by the fact that neonates (nursing on immune mothers) had significant (P less than 0.01) delayed footpad reactions to a crude T. spiralis antigen preparation, as compared with neonates nursing on non-immune controls.

The effects of ethanol consumption on the expression of immunity to Trichinella spiralis in rats.

The effects of ethanol ingestion on immune responses of female rats against Trichinella spiralis (T. spiralis) infections were investigated. Female rats were pair-fed either ethanol-containing or isocaloric control liquid diets for 68 days, during which time they underwent one pregnancy cycle. For some studies, animals were fed ethanol diets for 68 days beginning prior to pregnancy and continuing through lactation and involution. They were infected on Day 7 of involution with 1000 L1 larvae of T. spiralis and tested for a primary rejection response 10 days later. To test for a secondary immune response, rats were infected with T. spiralis, placed on ethanol-containing liquid diet 15 days later, and after 68 days on diets, challenged with 1000 T. spiralis larvae and killed 3 days later. For primary immunized studies, ethanol-treated animals demonstrated significantly lower levels of anti-T. spiralis serum antibodies in ELISA, reduced rates of H3 thymidine incorporation by lymph node cells stimulated with T. spiralis antigen and significantly higher numbers of intestinal worm burdens (decreased immunity) compared with pair-fed controls. For animals sensitized to T. spiralis prior to pregnancy and given a secondary challenge during involution, no differences were found between ethanol and pair-fed animals in their ability to reject their worm burdens or in anti-T. spiralis serum antibody levels; however, ethanol-treated animals showed reduced rates of thymidine incorporation by lymph node cells when stimulated with T. spiralis antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

Maternal to neonatal transmission of T-cell mediated immunity to Trichinella spiralis during lactation.

The potential of maternally derived cellular factors to mediate immunity to Trichinella spiralis in neonates during lactation was investigated in this study. Female FI rats, infected with T. spiralis, were able to transfer immunity to their suckling offspring, evidenced by a significant reduction in the intestinal parasite burdens of their neonates. When challenged between 2 and 3 weeks of age with 200 T. spiralis larvae, pups suckling on immune mothers harboured 28% and 26% (at 3 and 8 days post-challenge) of the worm numbers present in control neonates suckling on naive mothers. Cross-fostering experiments in which pups born of naive mothers but nursed by immune mothers showed significant immunity, demonstrated that this passage occurred through milk. The role of cell-mediated immunity in this immune transfer was analysed using T cells purified from MLN cells of syngeneic donor rats infected with T. spiralis. When 200 x 10(6) sensitized MLN T cells were adoptively transferred into lactating recipients, it led to the passive immunization of suckling neonates (26% and 13% of control values retained at 3 and 8 days post-challenge), while maternal injection of T cells primed to an irrelevant antigen (KLH) had no effect on neonatal immunity. Neonates fed per-orally with primed T lymphocytes early in lactation and prior to challenge were also rendered immune (34% and 44% of control values retained at 3 and 8 days post-challenge). A single dose of T. spiralis-primed T cells given to neonates in early lactation was sufficient to elicit a significant immune response in them at 2 weeks of age. These results support the hypothesis that cellular immunity mediated by antigen-specific T cells in milk can provide functional immune protection to the neonate against an intestinal pathogen.