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Effects of early feed restriction and cold temperature on lipid peroxidation, pulmonary vascular remodelling and ascites morbidity in broilers under normal and cold temperature.

Two experiments were conducted to evaluate the effects of early feed restriction on lipid peroxidation, pulmonary vascular remodelling and ascites incidence in broilers under normal and low ambient temperature. In experiment 1, the restricted birds were fed 8h per day either from 7 to 14 d or from 7 to 21 d, while the controlled birds were fed ad libitum. In experiment 2, the restricted birds were fed 80 or 60% of the previous 24-h feed consumption of full-fed controls for 7 d from 7 to 14 d. On d 14, half of the birds in each treatment both in experiment 1 and experiment 2 were exposed to low ambient temperature to induce ascites. Body weight and feed conversion ratio were measured weekly. The incidences of ascites and other disease were recorded to determine ascites morbidity and total mortality. Blood samples were taken on d 14, 21, 28, 35 and 42 to measure the plasma malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). On d 42, samples were taken to determine the right/total ventricular weight ratio (RV/TV), vessel wall area/vessel total area ratio (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA). Low-temperature treatment increased plasma MDA concentration. When broilers were exposed to a cool environment for 3 weeks, plasma SOD and GSH-Px activity were decreased compared with normal-temperature chicks. RV/TV, WA/TA and mMTPA on d 42 were increased in birds exposed to cold, consistent with the increased pulmonary hypertension and ascites morbidity. Early feed restriction markedly decreased plasma MDA concentration. The plasma SOD and GSH-Px activity of feed-restricted birds were markedly higher than those fed ad libitum on d 35 and d 42. All early feed restriction treatments reduced ascites morbidity and total mortality. On d 42, the RV/TV, WA/TA and mMTPA of feed-restricted broilers were lower than that of the ad libitum-fed broilers. The results suggested that early feed restriction alleviated the lipid peroxidation, promoted the activity of enzymatic antioxidant and inhibited pulmonary vascular remodelling. These changes might be associated with reduced ascites incidence.

Animals↗

Control of the development of the ovarian follicular hierarchy in broiler breeder pullets by food restriction during rearing.

1. The increase in the numbers of yellow and of white ovarian follicles was studied from 10 weeks of age to sexual maturity in broiler breeder pullets fed ad libitum and on a restricted intake. The growth of follicles greater than 1 mm diameter commenced at 14 to 16 weeks of age in ad libitum birds and at 22 to 24 weeks in those which were restricted. 2. In a second experiment, groups of birds restricted from 4 d of age were transferred to ad libitum feeding at 2-week intervals and examined at first egg. The numbers of yellow follicles at first egg were not affected by restriction to 14 weeks of age but, thereafter, declined linearly with time on restriction. 3. In the second experiment there were no comparable responses in the numbers of white follicles or in the rates of atresia. 4. Food restriction to 22 weeks of age produced about half the response in numbers of yellow follicles as restriction continued to point of lay (27 to 32 weeks of age). In practical situations, the results suggest that food intake should be restricted up to the onset of lay and then increased in relation to the rise in egg production. 5. Atresia was greatest in white follicles 3.15 to 6.30 mm diameter and was less outside this range.

Animals↗

Food restriction and development of thermoregulation in Muscovy ducklings (Cairina moschata).

1. The interaction between the effects of food restriction and cold stress on the development of body temperature, homeothermy index, metabolic rate and body weight were studied in Muscovy ducklings from hatching to 21 d of age. 2. The control group (ad libitum fed) and the food restricted group (fed to zero growth rate for 9 d) both became homeothermic when they were 2 d old with moderate (10 degrees C) cold stress. At severe cold stress (0 degrees C) the control group was homeothermic 5 days after hatching. However the food restricted group did not reach homeothermy at 0 degrees C and showed a large decrease (to hatching level) in homeothermy index at 10 degrees C and 0 degrees C after 9 d of food restriction. 3. Body temperature was lower in the food-restricted group during restriction and increased by 1 degree C after 24 h of ad libitum feeding. During food restriction, resting metabolic rate did not increase with age and was lower than the basal and existence metabolic rate predicted by Aschoff and Pohl (1970) and Kendeigh (1970) respectively. 4. The ratio of metabolisable energy (ME) intake to resting metabolic rate was 3 times lower in the food-restricted group than in the control group (0.09 and 0.27 respectively) on day 9. The availability of ME was more important than age for the development of thermoregulation in Muscovy ducklings. It is concluded that small improvements in the feeding regimen of young ducklings enhance the endurance and consequently reduce mortality from to environmental cold stress in a scavenger poultry system.

Aging↗

Welfare of food restricted male and female turkeys.

1. The welfare of male and female male-line turkeys fed ad libitum or food-restricted was determined at 4, 8, 12, 16, 20, 24, 28, 36(38) and 46(48) weeks of age using behavioural and physiological indices of well-being. Traditional turkeys fed ad libitum were kept as a control treatment. Restricted male and female male-line turkeys were fed to 0-5 during rearing and subsequently to 0-8 of sex-specific ad libitum-fed body weight. In another treatment, male-line males were fed ad libitum to 18 weeks and 0.8 of ad libitum thereafter. 2. Traditional turkeys and restricted male-line turkeys were more active than ad libitum-fed birds of both sexes. Restricted turkeys showed a high incidence of wall pecking. In the breeding period, about 0.4 of the observations of male-line males were of strutting behaviour whereas traditional male turkeys showed no strutting behaviour at the end of the breeding period. 3. The heterophil lymphocyte ratio (HLR) and the proportion of basophils were not increased in food-restricted turkeys. The HLR was relatively low in traditional birds, compared with male-line turkeys during the rearing period. 4. Plasma corticosterone concentrations were increased by food restriction during the rearing period. Corticosterone concentrations were relatively high in traditional turkeys at 4 and 8 weeks of age only. 5. Plasma lactate dehydrogenase (LIDH) activity was higher from 12 to 24 weeks of age in ad libitum-fed male-line turkeys and was consistent with mortality from cardiovascular disease in this group of turkeys. The pattern of activity of aspartate transaminase was similar, and alkaline phosphatase was inversely related to that of LDH. 6. It was concluded that turkeys may be better able to adjust physiologically to the demands of food restriction than broiler breeders and that there were few deleterious consequences of restricting male turkeys after 18 weeks of age. Male-line turkeys were less active than traditional turkeys.

Animal Feed↗

Small and growth-restricted babies: drawing the distinction.

UNLABELLED: The terms "small for gestational age" and "intrauterine growth restriction" have been used interchangeably to denote an in utero growth-restricted neonate. However, the two terms are not synonymous; not all small babies are growth restricted and not all growth-restricted ones are small. Research evidence, extending back to the middle of the last century, reveals that the number of growth-restricted babies who escape attention is not negligible and that the postnatal outcome of these babies is not uneventful. This paper highlights this issue and further discusses the available diagnostic tools for the identification of in utero-restricted neonates, that is, clinical assessment, anthropometric indices and obstetric ultrasound. Each of these tools has strengths and limitations, but, if combined, each could complement the other and help differentiate well-grown babies from those who are growth restricted. CONCLUSION: Identification of growth-restricted neonates is feasible through the integrated use of diagnostic tools.

Fetal Growth Retardation↗

Ovarian follicular dynamics in selected and control (relaxed selection) male- and female-lines of broiler breeders fed ad libitum or on restricted allocations of food.

1. The reproductive structures of 4 broiler breeder lines fed ad libitum or restricted were compared at 1st egg. The lines were contemporary male- and female-lines and their respective progenitor lines maintained without selection for 20 years. 2. The selected lines were larger and had more hierarchical yellow follicles than the relaxed lines. The numbers of positions in the hierarchy were greater in the selected lines. The number of atretic follicles and the proportion of multiple follicles were relatively high in the selected male-line. 3. There were more white follicles in the range 1.8 to 5.0 mm diameter in the selected male-line than in the relaxed line, whereas selection did not change white follicle numbers in the female-line. 4. Food restriction decreased the numbers of normal and atretic yellow follicles and the proportion of multiples but had no effect on the number of positions in the hierarchy. Restricted feeding decreased the number of white follicles 1.4 to 2.4 mm and increased follicles 2.4 to 5.0 mm diameter. 5. There was little change in age at 1st egg among lines fed ad libitum. Restricted feeding increased age at 1st egg and the difference was greater in female- than in male-lines and in relaxed compared with selected birds. 6. Abdominal fatness was similar among the lines fed ad libitum and was decreased by restricted feeding. 7. Oviduct weights were greater in selected than in relaxed lines and were decreased by restricted feeding only in the selected male-line. 8. The weight of the largest follicle was higher in the selected lines and in restricted compared with ad libitum-fed birds. Plasma triglyceride concentrations were lower in restricted birds from the selected and relaxed female-lines compared with the other groups. Differences in plasma oestrogen concentrations could not be detected.

Adipose Tissue↗

IgE class-restricted tolerance induced by neonatal administration of soluble or cell-bound IgE.

Induction of IgE class-restricted tolerance was studied in high IgE-responder (BALB/c X SJL)F1 mice, of which the parental BALB/c and SJL mice are high and low IgE-responder mice, respectively. 2,4-Dinitrophenyl (DNP)-specific monoclonal IgE was administered to (BALB/c X SJL)F1 mice neonatally in two forms: soluble IgE at 250 micrograms per injection, or 10-100 ng of IgE coupled to 25-50 X 10(6) syngeneic splenocytes by binding to the chemically reactive hapten trinitrobenzene sulfonate (TNBS) or directly conjugated via a heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). Polyclonal induction of IgE class-restricted tolerance was observed in (BALB/c X SJL)F1 mice, neonatally treated with soluble IgE or IgE-conjugated syngeneic splenocytes. Thus these mice failed to mount IgE antibody responses to either keyhole limpet hemocyanin or ovalbumin challenge, assessed by the passive cutaneous anaphylaxis reaction. The IgG antibody responses to these same antigens, however, were not affected by this treatment. The IgE class-restricted tolerance induced by both forms of IgE persisted up to at least 6 mo with repeated antigenic challenges. IgE coupled to syngeneic cells by TNBS or the SPDP method induced prolonged tolerance up to 9 mo. The induction of polyclonal IgE class-restricted tolerance was achieved only by monoclonal IgE, whereas DNP-specific monoclonal IgG1 plus IgG2b coupled to syngeneic splenocytes by the SPDP method failed to modulate either IgE or IgG antibody responses. In contrast, (BALB/c X A/J)F1 mice, of which both parental strains are high IgE responders, developed IgE class-restricted tolerance upon repeated neonatal injection of soluble IgE, but not IgE-conjugated syngeneic splenocytes, indicating that gene products of the low IgE-responder mice contributed to the effective presentation and/or recognition of epsilon heavy chain of the IgE molecules. Taken collectively, these results demonstrated that non-antigen-specific, isotype-restricted tolerance can be induced in the IgE antibody system. The differential induction of IgE class-restricted tolerance by different forms of tolerogen in the strains studied perhaps distinguishes two underlying cellular mechanisms for IgE class-restricted tolerance.

Animals↗

I-A-restricted T cell antigen recognition. Analysis of the roles of A alpha and A beta using DNA-mediated gene transfer.

The contributions of A alpha and A beta chains, and of subregions of A beta, to Ia-restricted recognition of antigen by Th lymphocytes were analyzed using a panel of L cells transfected with various pairs of A alpha b,d, or k genes and recombinant or wild-type A beta b,d, or k genes. The A beta genes included all possible exchanges of the whole NH2-terminal (beta 1) domain or halves of the beta 1 domain among these three allelic A beta genes. The Ia+ L cells derived from such transfections were used as antigen-presenting cells with a 21 member panel of responding Ia-restricted T hybridoma cells of differing nominal antigen specificity and Ia-restriction. Special care was taken to account for quantitative variation in levels of Ia expression throughout the experiments. The results of this analysis reveal that (a) only 2 of the 21 Th cells recognized Ia molecules involving either a nonparental A alpha or a nonparental A beta chain, and in both cases the degeneracy extended to only one of the two other alleles tested. This suggests that allele specific contributions from both A alpha and A beta chains are important in restricted recognition for most, if not all I-A-restricted Th cells. (b) In no case did substitution of the A beta 2 domain from either of the alternative haplotypes lead to any functionally detectable effects, demonstrating that polymorphisms in the A beta 1 domain can entirely account for the restriction imposed on Th cell responses by the entire A beta chain. (c) For 90% of the cells tested, replacement of the NH2-terminal portion of the beta 1 domain with an allogeneic segment led to Ia molecules unable to elicit Th responses. Furthermore, of all the cells permissive of the substitution of one or other half of the beta 1 domain, only two permitted the substitution of sequence from both alternative haplotypes. Taken together, these data strongly suggest that antigen recognition by most, if not all, I-A-restricted Th cells involves contributions from both halves of the A beta 1 domain. These data suggest that the role of I-A molecules in restricted Th cell recognition of antigen depends on conformational determinants unique to a particular combination of polymorphic alpha and beta chains, and that multiple such sites exist on a single Ia molecule.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

An endogenous processing pathway in vaccinia virus-infected cells for presentation of cytoplasmic antigens to class II-restricted T cells.

The recognition of virus-infected cells by class I MHC-restricted cytotoxic T cells requires endogenous processing of antigen for presentation. It is still unclear whether endogenous processing of antigen can be utilized by class II MHC molecules for presentation. To test this possibility, a human B cell line expressing HLA-A2 and HLA-DR1 was infected with a recombinant vaccinia virus expressing the Influenza A virus M1 matrix protein (VAC-M1) and was assayed for lysis by different M1-specific cytolytic T cell lines, restricted by either HLA-A2 or by HLA-DR1. Class II-restricted lysis of VAC-M1-infected cells did occur. This lysis required de novo M1 synthesis and was not due to exogenous antigen. Several properties of the endogenous processing pathway for class II-restricted presentation were different from those of the pathway utilized by class I molecules. First, class II-mediated recognition of VAC-M1 infected cells was less efficient, requiring higher doses of virus and longer infection times, than the class I-mediated recognition. Second, chloroquine completely blocked presentation of endogenous M1 to class II-restricted T cells but had no effect on the class I-restricted presentation. Third, the class II-restricted presentation of M1 was only mildly affected by Brefeldin A, a drug that prevents transport from the endoplasmic reticulum to the Golgi, whereas the class I-restricted presentation of M1 was completely abrogated by this drug. These data demonstrate the existence of an endogenous processing pathway for the presentation of cytosolic antigen by class II molecules and show that this pathway is distinct from the one used for presentation by class I molecules.

Anti-Bacterial Agents↗

Learning to overeat: maternal use of restrictive feeding practices promotes girls' eating in the absence of hunger.

BACKGROUND: Experimental findings causally link restrictive child-feeding practices to overeating in children. However, longitudinal data are needed to determine the extent to which restrictive feeding practices promote overeating. OBJECTIVES: Our objectives were to determine whether restrictive feeding practices foster girls' eating in the absence of hunger (EAH) and whether girls' weight status moderates the effects of restrictive feeding practices. DESIGN: Longitudinal data were used to create a study design featuring 2 maternal restriction factors (low and high), 2 weight-status factors (nonoverweight and overweight), and 3 time factors (ages 5, 7, and 9 y). RESULTS: Mean EAH increased significantly (P < 0.0001) from 5 to 9 y of age. Higher levels of restriction at 5 y of age predicted higher EAH at 7 y of age (P < 0.001) and at 9 y of age (P < 0.01). Girls who were already overweight at 5 y of age and who received higher levels of restriction had the highest EAH scores at 9 y of age (P < 0.05) and the greatest increases in EAH from 5 to 9 y of age (P < 0.01). CONCLUSIONS: The developmental increase in EAH from 5 to 9 y of age may be especially problematic in obesigenic environments. These longitudinal data provide evidence that maternal restriction can promote overeating. Girls who are already overweight at 5 y of age may be genetically predisposed to be especially responsive to environmental cues. These findings are not expected to be generalized to boys or to other racial and ethnic groups.

Adult↗

Suppression of benzo[a]pyrene metabolism by accumulation of triacylglycerols in rat hepatocytes: effect of high-fat and food-restricted diets.

Diet has been implicated as a major determinant of chemical carcinogenesis. Accordingly, rates of benzo[a]pyrene (B[a]P) metabolism were compared in hepatocytes isolated from rats maintained on control, high-fat or food-restricted AIN-76A diets. Rats maintained on the food-restricted diet were given 65% of food consumed by the control group fed ad libitum. The high-fat diet group had free access to a modified AIN-76A diet in which the amount of corn oil was increased 4-fold at the expense of digestible carbohydrates. The triacylglycerol content in hepatocytes varied in direct proportion to dietary fat and calories and was 66 +/- 5, 105 +/- 7 and 192 +/- 16 nmol/mg dry wt in cells isolated from rats fed food-restricted, control and high-fat diets respectively. In contrast, the rate of B[a]P metabolism was highest in hepatocytes from rats maintained on the food-restricted diet and lowest in cells from animals given the high-fat diet (i.e. food-restricted greater than control greater than high-fat). Thus, an inverse correlation existed between the rate of B[a]P metabolism and the content of triacylglycerols in hepatocytes. At a cell density of approximately 2 mg dry wt/ml, rates of B[a]P (40 microM) metabolism were 1324 +/- 186, 1150 +/- 198 and 829 +/- 76 pmol/mg dry wt/h, respectively, in hepatocytes isolated from rats fed food-restricted, control and high-fat diets. When cells were incubated with a lower concentration of B[a]P (10 microM), the rate of B[a]P metabolism was greater than 2-fold higher in hepatocytes from rats fed the food-restricted diet compared to the rate measured in cells from the high-fat group. Glucuronidation of B[a]P metabolites in hepatocytes from rats fed high-fat diet was also approximately 30% lower than rates determined for control and food-restricted groups. These diet-induced alterations in rates of B[a]P metabolism occurred in the absence of changes in specific activity of arylhydrocarbon hydroxylase or UDP-glucuronosyltransferase in liver microsomes. Further, the rate of 7-ethoxycoumarin metabolism, a more hydrophilic substrate, was not affected by diet and B[a]P but not 7-ethoxycoumarin accumulated in hepatic lipid droplets. Thus, diet-induced changes in intracellular triacylglycerol, particularly in lipid droplets, may alter access of B[a]P to binding sites on arylhydrocarbon hydroxylase and thereby modulate B[a]P metabolism in intact hepatocytes.

Analysis of Variance↗

Effect of age and food restriction on alkaline protease activity in rat liver.

The effect of age and food restriction on the hepatic alkaline protease activity of 100,000 x g supernatant has been investigated using 7-, 16-, and 26-month-old Fischer 344 rats. The proteasome, a major component of alkaline protease activity, is activated by sodium dodecyl sulfate (SDS) and this property was exploited to gain insight into the effects of age and food restriction on proteasome activity. Three alkaline protease activities, chymotrypsin-like (ChT-L), trypsin-like (T-L), and peptidylglutamyl peptide hydrolyzing (PGPH) activities were measured. These activities are also commonly used as measurement of proteasomal activities. Basal ChT-L and PGPH activities were not markedly altered by either age or food restriction. The level of T-L activity did not change with age, but was decreased by food restriction. SDS-activated ChT-L activity increased 15% between 7 and 26 months of age and this increase was blocked by food restriction. SDS-activated PGPH activity decreased 40% and the decrease was ameliorated by food restriction. In conclusion, we have shown that the alteration of alkaline protease activities by age and food restriction is not uniform and that the changes observed are likely due to alterations of proteasomal activity. The lack of uniformity in these alterations indicates that any assessment of alkaline protease activity requires the measurement of more than one of the enzymatic activities. Lastly, the first evidence suggesting that age and food restriction can modulate proteasomal activity is presented.

Aging↗

Alteration of rat liver 20S proteasome activities by age and food restriction.

The effects of age and food restriction on proteasome function in rat liver supernatant (100,000 x g) were investigated. The cellular level of the proteasome has been quantitated by using Western blot analysis. The level of the proteasome was not affected by either age or food restriction. The three best-characterized proteasomal peptidase activities, chymotrypsin-like (ChT-L), trypsin-like (T-L), and peptidylglutamyl peptide hydrolyzing (PGPH) activities, were measured in the presence and absence of the proteasomal activator, sodium dodecyl sulfate (SDS). Basal ChT-L, T-L, and PGPH activities were not markedly affected by either age or food restriction. SDS-stimulated ChT-L and T-L activities increased approximately 15% and approximately 30%, respectively, between 7 and 26 months of age, and the increase of both activities was prevented by food restriction. In marked contrast, the SDS-stimulated PGPH activity decreased approximately 40% with age. Food restriction, while not preventing the age-related decline, maintained higher levels of SDS-stimulated PGPH activity at all ages. The proteolytic activity of the proteasome toward casein was not altered by either age or food restriction. In conclusion, the cellular level of the proteasome as well as the caseinolytic activity of the proteasome appear to be unaffected by either age or food restriction. It appears unlikely that the proteasome activity changes are related to the reported age-associated decline of protein degradation. Simultaneously, proteasomal peptidase activities appear to be differentially regulated by both age and food restriction. It suggests more subtle age-related changes in proteasome function, which could include an effect on proteasomal subunit composition.

Aging↗

Effects of food restriction on mechanical properties of arterial system in adult and middle-aged rats.

The effects of food restriction on the mechanical properties of the vasculature were determined in Long-Evans male rats with different ages. Rats that began food restriction at the ages of 6 months and 12 months were fed on alternate days for 6 months. Rats at the ages of 12 and 18 months were referred to as adult and middle-aged rats and were anesthetized and thoracotomized. The exponentially tapered T-tube model was employed to relate pulsatile pressure and flow signals measured in the ascending aorta. In each age group, food restriction elicited a decrease in body weight as well as basal heart rate but showed no significant change in cardiac output. Arterial blood pressure, total peripheral resistance, and aortic characteristic impedance were not affected by food restriction in middle-aged rats. However, adult food-restricted rats exhibited lower mean arterial blood pressure (99.1 +/- 3.1 mmHg) than did adult ad libitum-fed rats (110.7 +/- 3.0 mmHg). Total peripheral resistance was reduced from 0.645 +/- 0.045 mmHg-min-kg/ml in adult ad libitum-fed rats to 0.492 +/- 0.030 mmHg-min-kg/ml in adult food-restricted rats. Moreover, aortic characteristic impedance of adult food-restricted rats (0.014 +/- 0.001 mmHg-min-kg/ml) was lower than that of adult ad libitum-fed rats (0.024 +/- 0.002 mmHg-min-kg/ml). Neither age nor diet exerted effects on wave transit time and produced no changes in aortic distensibility. In conclusion, food restriction may elicit significant changes in the mechanical properties of both Windkessel vessels and resistance arterioles in adult rats, but not in middle-aged rats.

Aging↗

The effect of dietary restriction of varying duration on survival, tumor patterns, immune function, and body temperature in B10C3F1 female mice.

Seven groups of mice were maintained on different dietary programs varying with respect to restriction at various stages of life. Restriction was associated with less age-related decline in T-dependent immunological function and a slight but significant lowering of body temperature. The best mean and maximum survival and the lowest late-life mortality rate was found in the group restricted throughout life, but restriction during any part of the lifespan enhanced survival to some degree. The mean life spans of tumor-bearing animals tended to be greater in restricted than in nonrestricted groups, corresponding to an age-decelerating effect. Tumor frequency varied with the period of life during which restriction took place and was not always decreased in restricted animals. These latter results suggest that the mechanisms whereby dietary restriction influences the aging rate and tumor susceptibility may not be entirely identical.

Animals↗

Influence of the restriction of individual dietary components on longevity and age-related disease of Fischer rats: the fat component and the mineral component.

The influence of restricting either the fat or the mineral component of the diet to the same extent as they are restricted in the life-prolonging, food-restriction paradigm but without restricting calories was studied in regard to longevity and age-related pathologic lesions of barrier-maintained male Fischer 344 rats. Neither the restriction of fat nor the restriction of mineral influenced the median length of life or maximum life span as indicated by the age of the 10th percentile survivors. Restricting the dietary fat did retard the development of chronic nephropathy and associated lesions, but it also increased the prevalence of lymphoma and leukemia. The development of chronic nephropathy was not significantly affected by restricting the mineral component of the diet.

Aging↗

The identification of tyrosine as a common key residue in unrelated H-2Kd restricted antigenic peptides.

We have compared the activity of several Kd- or Ld-restricted antigenic peptides as competitors in a functional competition assay using cytolytic T lymphocyte (CTL) clones. All of four unrelated Kd-restricted peptides tested could compete with each other but not with the Ld-restricted peptide P91A-. 12-24 (P91A). Moreover, the P91A peptide failed to compete with the four Kd-restricted peptides. In contrast, another Ld-restricted peptide [mouse cytomegalovirus (MCMV) pp89 167-176] could clearly compete with both Kd- and Ld-restricted peptides. The comparison of a series of modified MCMV pp89 peptides suggested that distinct structural features allow the interaction of the peptide with the two different MHC class I molecules. We showed previously that the competitor activity of two different Kd-restricted antigenic peptides was reduced substantially upon Ala substitution of the single Tyr residues present in these peptides. We now show a similar effect for two additional Kd-restricted peptides. Our results thus suggest that Tyr may function as an 'anchor' residue for many antigenic peptides that bind to the Kd molecule. Molecular modeling of the presumed antigen-binding site of the Kd molecule revealed the presence of two deep cavities that may be involved in binding peptide amino acid side chains. A model illustrating one possible interaction of a Tyr-containing peptide with the Kd molecule is presented.

Alanine↗

Comparison of HLA-DR1-restricted T cell response induced in HLA-DR1 transgenic mice deficient for murine MHC class II and HLA-DR1 transgenic mice expressing endogenous murine MHC class II molecules.

Transgenic mice expressing human HLA class II molecules provide a useful model for identifying HLA-restricted CD4+ epitopes. However, the influence of endogenous murine H-2-restricted T cell responses on HLA-restricted responses is not known. In the present study, we show that HLA-DR1 transgenic mice deficient for H-2 class II expression (HLA-DR1+/+/IAbeta0/0) exhibit an equivalent expression level of the transgene HLA-DR1 and a similar diversity in the TCR repertoire, but a slightly different number of CD4+ peripheral T cells, when compared to HLA-DR1 transgenic mice in which H-2 class II molecules were retained (HLA-DR1+/+/IAbeta+/+). More importantly, a strong antigen-specific HLA-DR1-restricted response was observed in nearly all HLA-DR1+/+/IAbeta0/0 mice immunized with HBV envelope protein (HBs) or capsid protein (HBc), whereas weak HBs- or HBc-specific HLA-DR1-restricted responses were detected in half of the immunized HLA-DR1+/+/IAbeta+/+ mice. Conversely, strong HBs- or HBc-specific H-2-restricted T cell responses were detected in HLA-DR1+/+/IAbeta+/+ mice but not in HLA-DR1+/+/IAbeta0/0 mice. Our results indicate that the coexpression of endogenous H-2 class II molecules reduces the intensity of HLA-DR1-restricted antigen-specific responses in transgenic mice, by favoring murine over human MHC recognition and education. Thus, HLA-DR1+/+/IAbeta0/0 mice represent a better model for identifying and characterizing HLA-DR1-restricted epitopes relevant for human disease.

Animals↗