Sheep Red Blood Cell Challenge Research Articles

An immune challenge of female great tits decreases offspring survival and has sex-specific effects on offspring body size

Investment in immunity is expected to decrease (costly immunity) or enhance (terminal investment) reproductive performance. Here, we tested the effects of activation of the immune system in female great tits (Parus major) on (1) their reproductive effort and (2) the survival and body condition of their offspring, controlling for chick sex. We injected females tending 3-day-old chicks with sheep red blood cells (SRBC) or saline (control) and recorded their provisioning rates 6 days later, during the expected peak of antibody production. We measured tarsus length and body mass in 11-day-old chicks and monitored changes in brood size. We found that female provisioning rates were unaffected by the SRBC challenge. An analysis without an outlier, however, showed a significant challenge-by-hatch date interaction. This interaction indicated that female provisioning rates decreased with hatch dates in the SRBC but not in the control nests, suggesting a stronger effect in later breeders. Chick body mass was not affected by female immunisation nor by its interaction with chick sex. However, we found a significant challenge-by-sex interaction on offspring tarsus. In SRBC nests, the difference in tarsus length between male and female chicks was lower than in controls, suggesting sex-dependent effects of the challenge on offspring structural growth. Finally, chick mortality was greater in SRBC nests compared with controls, but chick survival probability was not affected by sex. Overall, our results support the costly immunity but not the terminal investment hypothesis in the great tit.

Does paternal immunocompetence affect offspring vulnerability to maternal androgens? A study in domestic chickens.

ABSTRACTExposure of yolk androgens can positively stimulate chick growth and competitive ability, but may negatively affect immunity. It has been hypothesized that only chicks from immunologically superior fathers can bear the cost of prenatal exposure to high androgen levels. To test this hypothesis, we paired roosters from two selection lines, one up- and one down-selected for natural antibodies (NAbs), with hens from a control line. We measured yolk testosterone and androstenedione levels, and we injected the treatment group of eggs of each female with testosterone suspended in sesame oil and the control group with sesame oil only. We then measured hatching success and growth, and characterized the humoral and cellular immune responses using three different challenges: a phyto-hemagglutinin, a lipopolysaccharide and a sheep red blood cell challenge. We found that the hatching success, body mass, initial levels of natural antibodies and the chicks’ immunological responses to the three different challenges and development were affected neither by paternal immunocompetence nor by treatment. These results do not support the hypothesis that chicks from low-NAb line fathers are more sensitive to testosterone exposure during embryonic development than chicks from high-NAb line fathers.

The Impact of the Female Immune Challenge on Parental Feeding Rate, Mass and Body Condition Index of Offspring

Reproduction is a resource-demanding period, when trade-offs between investment in immunity versus other functions of an adult organism may exist. This research investigates the potential trade-off between a challenged immune system of a female parent and parental provisioning in free-living Great Tits Parus major and Eurasian Blue Tits Cyanistes caeruleus. In both species, half of females were challenged with sheep red blood cells (SRBC) as an antigen, while the others were injected with phosphate buffer saline (PBS) as a control. Six days later – at the day assumed to be the peak of an immune response in birds treated with SRBC, when nestlings were nine days old – parental feeding rates were recorded. Two days later (and eleven days after hatching), nestling mass and body condition index (BCI) were quantified. SRBC challenge of the female increased feeding rates of Great Tit males and females, but did not change parental provisioning in Eurasian Blue Tits. This may be explained by the different reproductive strategies in these two bird species. Parental provisioning influenced the mass and BCI of offspring. In both species, offspring mass was lower and BCI was higher in the SRBC group than those in the control group. This means that high providers, such as Great Tits with increased feeding rates, do not always raise chicks with high fledgling mass. Parental provisioning is one of the key factors linking reproduction and the immune system.

Liposome encapsulated all trans retinoic acid (ATRA) has enhanced immunomodulatory and inflammation reducing activities in mice model.

The all trans retinoic acid (ATRA) is found to have a promising regulatory effect on immune system and inflammatory responses in experimental research. The purpose of this study was to investigate whether this therapeutic efficiency of ATRA could be enhanced by encapsulating into a liposome formulation composed of Distearoyl-L-phosphatidylcholine (DSPC) and cholesterol utilizing a well-established mice model. The humoral antibody titer (HA), delayed-type hypersensitivity (DTH), bone marrow cellularity, hematology, and levels of α- esterase-positive cells, were taken as parameters to assess the level of immunomodulation in the sheep red blood cells (SRBC) immunized and challenged BALB/c mice. The anti-inflammatory effect of encapsulated ATRA was evaluated by the size changes in the induced inflammation edema in the mice paw as well as its histopathology. The results showed a significant immunostimulatory effect for both the free and encapsulated ATRA as indicated by the increase in the levels of total leukocyte, bone marrow and α-esterase positive cells and decreased Hb level respectively. We have also observed an enhanced specific antibody hemagglutinin titre value and the DTH response developed in response to SRBC challenge in these treatments. Both the immunostimulatory as well as inflammation reducing property were significantly higher in encapsulated ATRA treated group of mice over that of in free ATRA treated group of mice. Based on these results, we conclude that the encapsulated ATRA has a higher potency over free ATRA in its immunomodulatory activity and also has a significant impact on reducing inflammation in BALB/c mice model.

Estimation of crossbreeding parameters for humoral response in broiler

Crossbreeding parameters for humoral immune response – including the antibody production to sheep red blood cell challenge (SRBC) and serum haemolytic complement level (HC) – were estimated using a complete 4×4 diallel design with four synthetic broiler lines namely coloured synthetic male line (CM), white synthetic male line (WM), coloured synthetic female line (CF) and naked neck line (NN). The genetic group had a strong effect (P<0.01) on the mean SRBC and HC. Among 16 genetic groups, with 4 purebred and 12 crossbred groups, the crossbred progenies of NN×CM and CM×WM produced the highest mean SRBC (10.59±0.53) and HC (6.37±0.20), respectively. Estimates of direct line effect and maternal genetic effect for both SRBC and HC; average heterosis, direct line heterosis, and specific combining ability for HC were statistically significant (P<0.05). Most crossbred groups exhibited positive overall heterosis for both traits. Crossbred groups of WM×CM and CM×WM recorded the highest estimates of mean and percentage heterosis for SRBC (2.65 and 38.93%) and HC (1.72 and 36.84%), respectively. Results indicated that both additive and non-additive genetic variations were important for the humoral immune response in broiler. Differences in rankings of lines and crosses with respect to crossbreeding parameters suggested that genetic variations to these traits can be exploited by selection of specialised lines on the basis of response to SRBC challenge and serum HC level, or formation of a line based on optimum index values for both traits.

Diverse production system and social status effects on pig immunity and behavior

Pig performance, immunity, and behavior may be influenced by production system and social status. A conventional indoor housing system was compared with an outdoor system to determine the effects of diverse production systems on pig performance, behavior, and immunity. At 21 days of age, weight matched pigs reared in a conventional indoor or an outdoor production system were weaned and randomly selected for use in the present study. Six replicate pens were evaluated per treatment, with two littermate female pigs in each pen. Social status was determined at 7 weeks of age by a feed competition test. Pig behavior was recorded for 24 h using a scan sample technique. At 9 weeks of age, blood samples were taken from indoor- and outdoor-reared pigs and immune assays performed. Both indoor- and outdoor-reared pigs were given a sheep red blood cell (SRBC) challenge at 6 and 8 weeks of age, and antibody titers to SRBC were measured at 6, 7, and 9 weeks of age. Performance data were recorded throughout the study period. Body weight and average daily gain were not influenced by the production system in which the pig was reared. The production system effected standing, lying, drinking, and oral/nasal/facial behaviors. The production system influenced the expression of certain behaviors depending on the time of day. Immune and blood measures were affected by production system. The percentage of phagocytosis was greater (P ≤0.05) and antibody titers to SRBC challenge tended to be greater (P=0.066) among outdoor-reared pigs compared with indoor-reared pigs. Outdoor reared pigs had higher hemoglobin concentrations (P<0.005), percentage of hematocrit (P<0.005), mean corpuscular volume (P<0.005), and mean corpuscular hemoglobin (P<0.005) compared with indoor-reared pigs. Furthermore, dominance order influenced the immune system. Dominant pigs had greater phytohemagglutinin stimulated lymphocyte proliferation (P<0.01) and baseline antibody titers to sheep red blood cells (P<0.05) compared with submissive pigs. In conclusion, in the present study body weight and average daily gain were similar among diverse production systems. However, pig behavior and immune system were significantly influenced by the production system. Dominance order influenced the immune system regardless of the production system. © 2007 Elsevier B.V. All rights reserved.

Anti-inflammatory activity of cationic lipids.

1. The effect of liposome phospholipid composition has been assumed to be relatively unimportant because of the presumed inert nature of phospholipids. 2. We have previously shown that cationic liposome formulations used for gene therapy inhibit, through their cationic component, the synthesis by activated macrophages of the pro-inflammatory mediators nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha). 3. In this study, we have evaluated the ability of different cationic lipids to reduce footpad inflammation induced by carrageenan and by sheep red blood cell challenge. 4. Parenteral (i.p. or s.c) or local injection of the positively charged lipids dimethyldioctadecylammomium bromide (DDAB), dioleyoltrimethylammonium propane (DOTAP), dimyristoyltrimethylammonium propane (DMTAP) or dimethylaminoethanecarbamoyl cholesterol (DC-Chol) significantly reduced the inflammation observed in both models in a dose-dependent manner (maximum inhibition: 70-95%). 5. Cationic lipids associated with dioleyol- or dipalmitoyl-phosphatidylethanolamine retained their anti-inflammatory activity while cationic lipids associated with dipalmitoylphosphatidylcholine (DPPC) or dimyristoylphosphatidylglycerol (DMPG) showed no anti-inflammatory activity, indicating that the release of cationic lipids into the macrophage cytoplasm is a necessary step for anti-inflammatory activity. The anti-inflammatory activity of cationic lipids was abrogated by the addition of dipalmitoylphosphatidylethanolamine-poly(ethylene)glycol-2000 (DPPE-PEG2000) which blocks the interaction of cationic lipids with macrophages. 6. Because of the significant role of protein kinase C (PKC) in the inflammatory process we have determined whether the cationic lipids used in this study inhibit PKC activity. The cationic lipids significantly inhibited the activity of PKC but not the activity of a non-related protein kinase, PKA. The synthesis of interleukin-6 (IL-6), which is not dependent on PKC activity for its induction in macrophages, was not modified in vitro or in situ by cationic lipids. The synthesis of NO and TNF-alpha in macrophages, both of which are PKC-dependent, was downregulated by cationic lipids. 7. These results demonstrate that cationic lipids can be considered as novel anti-inflammatory agents. The downregulation of pro-inflammatory mediators through interaction of cationic lipids with the PKC pathway may explain this anti-inflammatory activity. Furthermore, since cationic lipids have intrinsic anti-inflammatory activity, cationic liposomes should be used with caution to deliver nucleic acids for gene therapy in vivo.

Effect of tumor necrosis factor-α on progesterone production by granulosa cells in laying hens of different genetic lines

In vitro progesterone production by granulosa cells in the presence or absence of human recombinant tumor necrosis factor-alpha (hrTNF-α) was measured at 10, 20, and 30 wk of egg production in White Leghorn hens selected for high (HA)- or low-antibody (LA) response to sheep red blood cell challenge. Isolated granulosa cells from the three largest preovulatory follicles (F1–F3) were incubated with 5 or 250 ng/ml hrTNF-α, and progesterone production was determined by the use of a validated radioimmunoassay. F1, F2, and F3 granulosa cells from HA hens produced more (P ⩽ 0.05) progesterone (140.8, 107.2, and 49.7 ng/ml) than LA hens (109.4, 78.9, and 26.9 ng/ml). The treatment of granulosa cells with hrTNF-α consistently inhibited (P ⩽ 0.05) progesterone secretion by all follicles among HA and LA hens, but not always at both doses. Generally, 5 ng/ml hrTNF-α was the maximum inhibitory dose. In the laying hen, a decrease in steroid production in response to cytokines may upset the steroid balance created by follicular hierarchy and inhibit or delay ovulation.

A Nucleoside-Nucleotide Mixture and Its Components Increase Lymphoproliferative and Delayed Hypersensitivity Responses in Mice

We studied the effect of individual components of a nucleoside-nucleotide mixture on T cell-mediated immunity in BALB/c and DBA/2 mice. Mice were fed for 4 wk a nucleotide-free 20% casein diet (control) or this diet supplemented with 3 mol/kg of one of the following; inosine, guanosine monophosphate, uridine, cytidine, thymidine, or a mixture of these. In both strains of mice, popliteal lymph node immunoproliferative response to alloantigeneic (C57BL/6 splenic cells) challenge in mice fed the mixture and guanosine monophosphate was greater (P < 0.05) than in the mice fed the control diet. BALB/c mice fed inosine, uridine, and thymidine also showed greater (P < 0.05) responses compared with control fed mice. In the sheep red blood cells challenge assay, foot pad weight-gain in both strains of mice fed the mixture and the individual components supplemented diets was greater (P < 0.05) than in those fed the control diet. In both strains of mice, interleukin-2 secretion by popliteal lymph node lymphocytes in mice fed the mixture and thymidine was higher (P < 0.05) compared with the rest of the groups except BALB/c mice fed cytidine. Significantly higher than control secretions of interferon-γ were observed only in BALB/c mice fed inosine, thymidine and the mixture. We conclude that mice fed the nucleoside-nucleotide mixture and the individual components of the mixture had greater responses in the T cell-mediated immune functions studied and that the responses in mice fed the mixture were not different from those in mice fed the individual components.

Antigen-induced alveolitis: Cytokine production in a mouse model

Mice of the C57BL/6 strain were injected intraperitoneally with 10(8) sheep red blood cells (SRBC), then instilled intratracheally with 10(8) SRBC two to three weeks later. After a single intratracheal exposure, a significant cellular infiltrate occurred, composed mostly of macrophages and lymphocytes. Lymphocytes proliferated significantly in response to SRBC antigen in vitro and released interleukin-2 (IL-2). Alveolar macrophages isolated from mice challenged with SRBC released higher levels of IL-1, IL-6, and tumor necrosis factor-alpha (TNF-alpha) upon in vitro lipopolysaccharide (LPS) stimulation compared to unprimed, challenged mice or mice receiving intraperitoneal SRBC alone. Lymphocytes from primed mice challenged three times with SRBC proliferated significantly less in response to the antigen than mice receiving one SRBC challenge and released significant levels of interferon gamma (IFN-gamma). Bronchoalveolar macrophages isolated from primed mice given three SRBC challenges released slightly higher levels of TNF-alpha and IL-6 in response to LPS than those from unprimed mice. After the third instillation, levels of hydroxyproline in the lungs increased significantly, indicative of a fibrotic reaction. Neutralization of IL-1 (by anti-mouse type 1 IL-1 receptor) or TNF-alpha resulted in the partial abrogation of the initial neutrophil influx, with some effect on the subsequent lymphocyte and macrophage influx. Blocking IL-1 or IL-2 but not TNF-alpha also resulted in a significant decrease in lung hydroxyproline increase, as well as lung granulomatous response and fibrosis. Overall, these results suggest that lymphoproliferation in the lungs in response to an antigen can result in fibrosis, mediated in part by IL-2 and IL-1.

Acute Inflammatory Response to Sheep Red Blood Cell Challenge in Mice Treated with 2,3,7,8-Tetrachlorodibenzo- p-dioxin (TCDD): Phenotypic and Functional Analysis of Peritoneal Exudate Cells

TCDD is a widespread environmental contaminant of concern to human health because of its well-recognized immunotoxicity in laboratory animals. Suppression of the murine antibody response to xenogeneic erythrocytes has been shown to be one of the most sensitive assays for TCDD immunotoxicity. However, the cellular mechanisms underlying the suppressed immune function have not been fully elucidated. In the present studies, peritoneal macrophage recruitment, activation, and antigen-presenting function in response to sheep red blood cell (SRBC) injection were compared in C57B1/6 mice treated with a single oral dose of 0 or 5 μg TCDD/kg. In vehicle-treated mice, SRBC injection induced a typical inflammatory response in the peritoneal cavity. Within 6 hr, the number of neutrophils increased and remained elevated until 40 hr. Macrophage numbers increased at 24 hr and remained elevated through 72 hr. In TCDD-treated mice, a hyperinflammatory response to SRBC was observed. The total number of peritoneal exudate cells was significantly greater at 16, 24, and 40 hr after SRBC challenge when compared to that of vehicle-treated mice. The increased number of peritoneal cells reflected significant increases in both neutrophils and macrophages. Mac-1 + peritoneal cells were examined by two-color flow cytometric analysis on Days 0-3 after SRBC injection for expression of the activation markers F4/80 and I-A. The intensity of F4/80 fluorescence significantly decreased 24-72 hr following SRBC challenge, while fluorescence associated with I-A significantly increased at 72 hr. These changes are consistent with macrophage activation. TCDD did not significantly alter F4180 expression on Mac-1 + cells, whereas I-A expression was increased earlier on cells from TCDD-treated mice. However, TCDD treatment did not alter the antigen presentation function of peritoneal cells, assessed by their ability to induce the proliferation of SRBC-primed T cells in vitro. The antigen-presenting function of adherent spleen cells was also not altered by TCDD exposure. To test the hypothesis that an excess number of phagocytes in TCDD-treated mice were clearing the antigen more efficiently, leading to a smaller (e.g., suppressed) antibody response, we attempted to overcome TCDD suppression by increasing the amount of SRBC antigen used for challenge. However, the magnitude of the anti-SRBC response in TCDD-treated mice was not significantly altered by increasing the antigen challenge dose, suggesting that enhanced clearance of antigen by macrophage is not a mechanism for TCDD-induced suppression of the anti-SRBC response. The mechanism(s) for the enhanced inflammatory response and its relationship, if any, to TCDD immunotoxicity remain to be elucidated.

The heterogeneity of the age-related decline in immune response: Impairment in delayed-type hypersensitivity and cytotoxic T-lymphocyte activity occur independently

Aged mice are known to have a heterogeneous cytotoxic T-lymphocyte (CTL) response to an influenza challenge. We sought to determine whether delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) was also heterogeneous in aging and, if so, whether this correlated with anti-influenza CTL activity. Aged mice developed mean footpad swelling of 0.24 ± 0.09 mm following SRBC challenge and mean CTL activity of 35 ± 18%. Even though these values were significantly lower than those from young mice (0.54 ± 0.10 mm and 51 ± 2%, respectively), no correlation was found between anti-influenza CTL activity and DTH to SRBC ( r = 0.01). We believe the most likely reason for this independent decline in immune response is because different activation signals are required for DTH and CTL activity. Our data thus suggest an age-related functional mosaicism in the immune system.

EFFECTS OF INGESTED LEAD ON ANTIBODY PRODUCTION IN MALLARDS (ANAS PLATYRHYNCHOS)

We examined the effects of lead ingestion on in vitro and in vivo indices of immune function in mallards (Anas platyrhynchos). Twenty-four mallard drakes were randomly divided into three groups (I, II, III), then assigned to treatment or control subgroups (n = 4). On day 0, all treatment birds were dosed orally with one number 4 lead shot (mean = 0.2 g). We challenged all individuals in each group with washed sheep red blood cells (SRBC) injected intraperitoneally on days 0 (Group I), 7 (Group II) or 14 (Group III), and collected blood for analyses 7 and 8 days after SRBC challenge. We measured and compared blood lead concentrations, in vitro lymphocyte transformation responses to phytohemagglutinin A and lipopolysaccharide, and hemagglutination titers to SRBC. Mean blood lead concentrations were elevated (P less than or equal to 0.04) in treatment birds at each sampling period. Large individual variability in lymphocyte stimulation responses precluded further analysis of those data. Hemagglutination titers to SRBC were lower (P less than 0.0001) in lead-poisoned ducks than in controls, suggesting that ingested lead may have immunosuppressive effects on mallards.

Effects of verrucarin A and roridin A, macrocyclic trichothecene mycotoxins, on the murine immune system

Verrucarin A (Ver A) and roridin A (Ror A), macrocyclic trichothecene mycotoxins, were examined for their immunomodulatory effects. Both mycotoxins were administered intraperitoneally at 0.35 mg/kg (1/2 the LD50) in CD-1 mice. Lymphocyte proliferation was studied on days 2, 4 and 7 after animals were dosed with Ver A. After day 2, no significant differences in [3H]thymidine (3H-TdR) incorporation were observed using concanavalin A (Con A), phytohemagglutinin (PHA), pokeweed mitogen (PWM) or lipopolysaccharide (LPS). On day 4 DNA synthesis induced by Con A, PHA and PWM was increased significantly. On day 7, PHA stimulation increased (p less than 0.001) above controls while Con A, PWM and LPS responses were not significantly different. The data indicated that Con A, PHA and PWM responses were time-dependent. Antibody production was evaluated by the hemolytic plaque assay; sheep red blood cells (SRBC) and Ver A were administered simultaneously, and Ver A was given 2 days after SRBC challenge. While spleen weights increased when Ver A was administered with SRBC or when the toxin was given 2 days after antigen, the antibody responses were not altered. In contrast, roridin A decreased PHA stimulation only on day 7 (p less than 0.05). There were no significant effects associated with the other mitogens. Antibody production did not differ significantly following administration of Ror A. Although Ver A and Ror A are equitoxic and structurally similar, their immunomodulatory properties for the given dose differ considerably. These different immunologic responses may be independent of other systemic toxic effects.

Effect of aging on immunological memory in gastrointestinal tract induced by sheep red blood cells in mice.

Tolerance induction by oral administration of sheep red blood cells (SRBC) was investigated in young and aged mice. Two-month old C3H/He mice receiving oral administration of 8 X 10(9) SRBC a day for two weeks became tolerant to a subsequent SRBC challenge in the systemic immune system. In contrast, older C3H/He mice aged about 1 year old receiving the same treatment did not become tolerant but resisted tolerance induction and produced a prominent IgG memory in the systemic immune system. Autoimmune-prone NZB mice showed a similar resistance to tolerance induction even at 3 months of age. To investigate a possible role of the liver in the oral tolerance induction, young (2-month old) C3H/He mice received an injection of 1 X 10(8) SRBC via portal vein. The mice thus treated, were not tolerized at all but immunized. The results suggest that gut-associated local immune system play a key role in the induction of the oral tolerance and that the tolerance inducing function of the local immune system declines with aging. Antigens in the gut including denatured self antigens may immunize the systemic immune system of aged animals.

Murine bone marrow IgA responses to orally administered sheep erythrocytes.

Specific immunization protocols have been established for the induction of murine bone marrow IgA responses to the T cell-dependent (TD) antigen sheep red blood cells (SRBC). Systemic immunization, either i.p. or i.v., followed by a second injection, induced splenic IgM and IgG responses and a bone marrow IgM response. No significant IgA responses were observed in either lymphoid tissue compartment. Oral immunization with SRBC by gastric intubation for 2 days, followed 1 wk later by an i.p. injection of SRBC resulted in a splenic IgA plaque-forming cell (PFC) response, but did not elicit a bone marrow IgA response. Repeated daily gastric intubation of SRBC to C3H/HeN and C3H/HeJ mice led to the previously reported pattern of systemic unresponsiveness in C3H/HeN mice and good anamnestic type IgM, IgG, and IgA splenic anti-SRBC PFC responses in the C3H/HeJ strain upon parenteral challenge. Oral administration of SRBC for 14 days to C3H/HeN mice, followed by systemic SRBC challenge, resulted in diminished splenic PFC responses of all isotypes, whereas gastric intubation of SRBC for 28 days led to complete systemic unresponsiveness to antigen in C3H/HeN mice. Interestingly, the repeated oral administration of SRBC resulted in significant bone marrow IgA PFC responses upon i.p. challenge in both C3H/HeN and C3H/HeJ mouse strains. The bone marrow IgA responses were clearly dependent upon chronic oral exposure to SRBC, because gastric intubation with SRBC for 2 consecutive days/wk for 10 wk also induced bone marrow and splenic IgA anti-SRBC PFC responses in C3H/HeN mice. These results suggest that memory B cells reside in the bone marrow of orally immunized mice and can yield anamnestic-type responses to challenge with the inducing antigen. The memory cells may arise in the Peyer's patches of the gut and migrate to the bone marrow. The possibility that the bone marrow is a component of the common mucosal immune system in mammals is suggested by this study.

Changes in plasma, tissue, and urinary nitrogen metabolites due to an inflammatory challenge.

Studies were undertaken to define the changes in protein metabolism that result from stimulation of the immune system by noninfectious inflammatory agents. Chicks were injected with inflammatory agents and metabolite concentrations were determined between 4 and 48 hr postchallenge. Inflammatory agents resulted in a generalized decrease in the concentration of plasma nitrogen metabolites, including ammonia, uric acid, urea, and several amino acids. Escherichia coli and sheep red blood cell (SRBC) injections induced changes in the concentrations of tissue-free amino acids at 16 hr postchallenge. After E. coli injections, free amino acid concentrations were increased by 175% in muscle and decreased by approximately 25% in liver, spleen, and bursa. A SRBC challenge resulted in similar decreases in free amino acid concentrations in the spleen and bursa as did E. coli; however, muscle and liver free amino acid concentrations were mostly unchanged. Urinary ammonia was increased, urinary uric acid was decreased, and urinary amino acids were not affected by E. coli injection. These findings indicate that stimulation of the immune system by noninfectious inflammatory agents induces tissue-specific changes in nitrogen metabolism. Changes in amino acid pool sizes in various tissues suggest alterations in rates of protein synthesis or degradation.

Collaboration between thymus-derived and marrow-derived thoracic duct lymphocytes in the hemolysin response of the rat

Thymectomized (Tx), irradiated hooded rats, reconstituted with 200 × 10 6 syngeneic normal bone marrow cells, showed variable responsiveness to challenge with sheep red blood cells (SRBC) at four and eight weeks after irradiation. If Tx recipients were restored instead with only 10 × 10 6 marrow cells from lymphocyte-depleted (i.e. thymectomized, thoracic duct-drained) donors, these rats were unresponsive to SRBC challenge at four weeks. Injection of 200 × 10 6 syngeneic thymus cells into such rats restored the hemolysin response to SRBC and the antibody was made by marrow-derived cells. Thoracic duct lymphocytes obtained from Tx irradiated rats reconstituted with lymphocyte-depleted marrow were able to transfer responsiveness to this antigen to irradiated secondary recipients if thymocytes were injected one week or more before cannulation. The restoration of competence in the thoracic duct population required the presence of thymus-derived cells but all antibody made in secondary recipients was synthesized by marrow-derived cells. It is argued that the competence of normal thoracic duct lymphocytes is due to the presence of a mixture of thymus-derived and marrow-derived cells.