Genetic control of immune responses of inbred mice: Responses against terpolymers poly(glu57lys38ala5) and poly(glu54lys36ala10)

The immune response patterns of inbred and congenic strains of mice against terpolymers poly(glu⁵⁷lys³⁸ala⁵) and poly(glu⁵⁴lys³⁶ala¹⁰) have been studied. Initial recognition of the polymers is ascribed to ‘GA’ receptors (Ir-GA gene product) on T cells of mice ofH-2 haplotypes,a,b,f,k ands, and ‘GL’ receptors (Ir-GL gene product) of mice ofH-2 ^p,H-2 ^q andH-2 ^j haplotypes, and to GA and/or GL receptors of mice ofH- 2^d andH- 2^r haplotypes. The specificity of the antibody is directed predominantly against GL. The inability to elicit antibody with GA specificity has been ascribed to the lack of significant concentrations of GA sequences in the polymers to interact with appropriate receptors on B cells. The weakest responders were mice of H-2^b haplotype. F¹ hybrids (responders×nonresponders) were all responders demonstrating the dominant character of responsiveness. Wide variations in antibody levels produced among strains of mice of theH-2 ^k andH-2 ^b haplotypes are ascribed to genes not linked toH-2.     

Multigenic control of immune responses of inbred mice against the terpolymers poly(Glu57Lys38Ala5) and poly(Glu54Lys36Ala10) and linkage withH-2 haplotype

Results of immunizations of recombinant inbred and congenic strains of mice with the random polymers poly(glu⁵⁷ lys³⁸ala⁵) or GLA⁵ and poly(glu⁵⁴lys³⁶ala¹⁰) or GLA¹⁰ indicate that there is an association of the responsiveness with theH-2 haplotype. Although the C57BL/6J mice (H-2 ^b haplotype) are “non responders”, the C57BL/6By originally derived from mice of the same haplotype are responders. The immune response pattern of recombinant strains carrying haplotypes derived by crossing over within theH-2 complex indicate that the responsiveness is under control of anIr gene which maps to the left of theIB subregion. Studies with the backcross mice indicated multigenic control of the responsiveness, with one locus beingH-2 linked and another locus segregating independently ofH-2.     

Genetic control of T-cell proliferative responses to poly(glu40ala60) and poly(glu51lys34tyr15): Subregion-specific inhibition of the responses with monoclonal Ia antibodies

The relationship betweenIr genes and Ia antigens was studied in the T-cell proliferative responses to two synthetic polypeptides poly(glu⁴⁰ala⁶⁰) (GA) and poly(glu⁵¹lys³⁴tyr¹⁵) (GLT¹⁵). The response to GA was found to be controlled by anIr gene in theI-A subregion, whereas the anti-GLT¹⁵ response was shown to be under dual control, oneIr gene mapping probably in theI-A subregion, and the other in theI-E subregion. We obtained two different lines of evidence suggesting identity ofIr and Ia genes. First, the presence of certain serologically identified allelic forms of the I-A-encoded A molecule correlated with the responder status to GA both in inbred strains and in B10.W lines, the latter carrying wild-derivedH-2 haplotypes. Thus the Ir and Ia phenotypes were not separable in strains of independent origin. Second, the anti-GA response was completely inhibited by monoclonal antibodies against determinants on the A molecule (Ia.8, 15, and 19), but not by a monoclonal antibody against a determinant on the E molecule (Ia.7). In contrast, the anti-GLT¹⁵ response was only inhibited by a monoclonal antibody against the E molecule, but not by antibodies against the A molecule. Our data support the hypothesis that Ia antigens, as restriction elements for T-cell recognition, may in fact be the phenotypic manifestation ofIr genes.     

Immune responses of mice against poly(glu lys ala) terpolymers and linkage with H-2

The immune responses of inbred mice to the terpolymers poly(glu⁴⁸-Iys³² ala²⁰) GLA²⁰, poly(glu³⁶lys²⁴ala⁴⁰) GLA⁴⁰, and poly(glu²⁴lys¹⁶ala⁶⁰) GLA⁶⁰ were studied. Antibody levels were measured with the homologous, as well as with the crossreacting polymers (glu⁶⁰ala⁴⁰) GA and (glu⁶⁰lys⁴⁰) GL. It was determined that the terpolymers consist of many determinants of varying immunogenic strengths which account for the dose dependency requirements for responsiveness as follows: mice ofH-2 haplotypesa, b, d, k, ands respond to ten and 100g GLA²⁰ and GLA⁴⁰ and to one, ten, and 100 g GLA⁶⁰; mice ofH-2 haplotypesp, q, andr do not respond well to any concentration of GLA²⁰ but respond well to 100 g GLA⁴⁰ and teng GLA⁶⁰. That the congenic mice C3H.NB (H-2 ^p) and B10.R111 (H-2 ^r), having responder backgrounds of C3H (H-2 ^k) and C57BL/10 (H-2 ^b) mice, respectively, do not respond would suggest strongly that there is linkage of responsiveness toH-2 in the above strains. In addition, the responsiveness of AQR mice to GLA⁶⁰ would map theIr gene(s) to the right of theK region, and most likely in theI region. The antibody against GLA²⁰ was directed against GL. Responses of miceof H-2 haplotypesp, q, andr against GLA⁴⁰ and GLA⁶⁰ were directed predominantly against unique GLA determinants that were neither GA nor GL. Mice of the other respondingH-2 haplotypes (a, b, d, k, ands) produced antibody against these unique GLA specificities, as well as against GL and/or GA determinants. The importance of measuring responses with the homologous polymer is therefore demonstrated. It was postulated that the recognition of GLA²⁰ at the T-cell level is via GLA determinants having a limited amount of alanine, which are different from those helical GLA determinants recognized in the polymers GLA⁴⁰ and GLA⁶⁰.     

Mechanisms of genetic control of immune responses

We examined multiple genetically regulated Immoral and cell-mediated immune (CMI) responses to poly(glu⁶⁰ala³⁰tyr¹⁰) (GAT) using a panel of mouse strains. We show that assignment of responder/nonresponder status depends upon the assay method. In addition, two distinct categories of nonresponder mice were found: (1) those which are unresponsive by all parameters tested (H-2 ^q and H-2 ^s haplotypes) and (2) those which are partially nonresponsive [H-2 ^bm12 mutant strain—a low/nonresponder by splenic plaque-forming cell (PFC) and delayed-type hypersensitivity (DTH) responses, but exhibits B6 parental levels of high GAT-specific T-cell proliferation (Tprlf) and interleukin-2 production]. The distinction between these two nonresponder types was confirmed by complementation tests in which significant GAT-specific PFC and DTH responses were seen in (H-2 ^q × H-2 ^bm12)F₁ hybrids, but not in (H-2 ^q × H-2 ^s )F₁ hybrids. Suppressor T cells (Ts) also play a selective role in nonresponsiveness to GAT. Cyclophosphamide treatment of nonresponders (to eliminate Ts activity) as well as immunization with GAT coupled to the immunogenic carrier MBSA result in the development of GAT-specific humoral, but not CMI responses. Our results indicate that the T cell is the cellular site of Ir gene expression and that Tprlf responses do not correlate with functional helper T-cell activity and suggest distinct, multi-step Th/Ts regulatory pathways in the development of humoral and CMI effector functions.     

Regulation of the Na+-coupled glutamate transporter EAAT3 by PIKfyve

The Na⁺,glutamate cotransporter EAAT3 is expressed in a wide variety of tissues. It accomplishes transepithelial transport and the cellular uptake of acidic amino acids. Regulation of EAAT3 activity involves a signaling cascade including the phosphatidylinositol-3 (PI3)-kinase, the phosphoinositide dependent kinase PDK1, and the serum and glucocorticoid inducible kinase SGK1. Targets of SGK1 include the mammalian phosphatidylinositol-3-phosphate-5-kinase PIKfyve (PIP5K3). The present experiments explored whether PIKfyve participates in the regulation of EAAT3 activity. To this end, EAAT3 was expressed in Xenopus oocytes with or without SGK1 and/or PIKfyve and glutamate-induced current (I_glu) determined by dual electrode voltage clamp. In Xenopus oocytes expressing EAAT3 but not in water injected oocytes glutamate induced an inwardly directed I_glu. Coexpression of either, SGK1 or PIKfyve, significantly enhanced I_glu in EAAT3 expressing oocytes. The increased I_glu was paralleled by increased EAAT3 protein abundance in the oocyte cell membrane. I_glu and EAAT3 protein abundance were significantly larger in oocytes coexpressing EAAT3, SGK1 and PIKfyve than in oocytes expressing EAAT3 and either, SGK1 or PIKfyve, alone. Coexpression of the inactive SGK1 mutant ^K127NSGK1 did not significantly alter I_glu in EAAT3 expressing oocytes and completely reversed the stimulating effect of PIKfyve coexpression on I_glu. The stimulating effect of PIKfyve on I_glu was abolished by replacement of the serine by alanine in the SGK consensus sequence (^S318APIKfyve). Moreover, additional coexpression of ^S318APIKfyve significantly blunted I_glu in Xenopus oocytes coexpressing SGK1 and EAAT3. The observations demonstrate that PIKfyve participates in EAAT3 regulation likely downstream of SGK1.     

Murine responses to (Tyr-Glu-Ala-Gly)n: II. H-2 and non-H-2 gene effects

Mice of the H-2^b haplotype responded to the sequential polymer poly(Tyr-Glu-Ala-Gly) in the in vitro T-cell proliferative assay, irrespective of whether they were homozygous or heterozygous at the H-2^b locus. The antibody responses of the H-2^b congenic mice to this polymer were variable, with A.BY and BALB.B showing responses better than those of C57BL/6 and C57BL/10 strains. The antibody responses of the F₁ progeny of (responder × nonresponder) strains of mice to this polymer are generally lower than the responder parents. F₁ mice with C57BL/10 background were the poorest responders. Studies with F₂ mice and backcross progenies of selective breeding of high and low antibody responder (C57BL/6 × BALB/c) F₁ to high responder C57BL/6 mice indicated that both non-H-2 genes and H-2 gene dosage effects influenced the magnitude of the humoral antibody responses. Animals having low responder non-H-2 background and only half the dosage of the responder immune response genes has greatly diminished antibody responses.     

Recombination between genes coding for immune response and the serologically determined antigens in the chickenB system

Evidence is presented for a crossover between the genes coding for the serologically determined (SD) antigens on erythrocytes and an immune response gene (Ir-GAT) controlling immune response to the synthetic polypeptide GAT within theB complex, the MHC of chickens. TheIr-GAT ¹ andIr-GAT ¹⁹ alleles control low and high immune response to GAT, respectively. Both low and high responders were recovered as recombinants fromB ¹ B ¹ andB ¹⁹ B ¹⁹ birds. The low-responder haplotypes are homozygous for theIr-GAT ¹ allele and the high-responder haplotypes carry theIr-GAT ¹⁹ allele. Mortality forB ¹ B ¹ nonresponder birds was 39%, compared with 19% for theB ¹ B ¹ high responders; this suggests the possibility that genes located within the immune response region of theB complex exert some genetic control over viability and survival.The following abbreviations are used in this paper MHC major histocompatibility complex - Ir immune response - SD serologically determined - GA (L-glutamic acid⁶⁰, L-alanine⁴⁰) n - DNP-GL dinitrophenyl-(L-glutamic acid⁶⁰, L-lysine⁴⁰) - PLL poly-L-lysine - (T,G)-A--L poly-(L-tyrosine-L-glutamic acid)-poly-D, L-alanine-poly-L-lysine - GAT, GAT¹⁰ (L-glutamic acid⁶⁰ L-alanine³⁰ L-tyrosine¹⁰) n - CFA complete Freund's adjuvant - PBS phosphate-buffered saline     

Absolute stereochemistry of leucinopine,a crown gall opine

Crown gall tumors incited by Agrobacterium tumefaciens strain Bo542 have been reported to synthesize a tumor-specific substance identified as N-(1,3-dicarboxypropyl)-leucine (leucinopine), a compound with two centers of asymmetry. We report here evidence that leucinopine is indeed a crown gall opine, in that it is specifically catabolized by A. tumefaciens strains carrying the tumor-inducing plasmid pTi Bo542, as well as strains carrying closely related plasmids pTi AT1 and pTi AT4. We further report catabolism of leucinopine by the succinamopine-type strains A518, A519 and A532, carrying pTi EU6, pTi AT181 and pTi T10/73, respectively. Strains lacking any virulence plasmid, as well as those carrying octopine or nopaline type Ti plasmids or mannopine type Ri plasmids, did not catabolize leucinopine. On the basis of specificity of catabolism by bacteria carrying pTi Bo542, we conclude that the stereochemistry of natural leucinopine is l-threo, i.e. l^glu,l^leu. Such stereochemistry is novel in the opines known thus far: octopine, nopaline and succinamopine have d,l-stereochemistry: d^ala,l^arg (octopine), d^glu,l^arg (nopaline) and d^glu,l^asn (succinamopine).     

Development and Partial Characterization of Nearly Isogenic Pea Lines (Pisum sativum L.) that Alter Uptake Hydrogenase Activity in Symbiotic Rhizobium

Some Rhizobium bacteria have H₂-uptake (Hup) systems that oxidize H₂ evolved from nitrogenase in leguminous root nodules. Pea (Pisum sativum L.) cultivars `JI1205' and `Alaska' produce high Hup (Hup⁺⁺) and moderate Hup (Hup⁺) phenotypes, respectively, in Rhizobium leguminosarum 128C53. The physiological significance and biochemical basis of this host plant genetic effect are unknown. The purpose of this investigation was to advance basic Hup studies by developing nearly isogenic lines of peas that alter Hup phenotypes in R. leguminosarum strains containing hup genes. Eight pairs of nearly isogenic pea lines that produce Hup⁺⁺ and Hup⁺ phenotypes in R. leguminosarum 128C53 were identified in 173 F₂-derived F₆ families produced from crosses between JI1205 and Alaska. Tests with the pea isolines and three strains of hup-containing R. leguminosarum showed that the isolines altered Hup activity significantly (P ≤ 0.05) in 19 of 24 symbiotic combinations. Analyses of Hup phenotypes in F₆ families, the F₁ population, and two backcrosses suggested involvement of a single genetic locus. Three of the eight pairs of isolines were identified as being suitable for physiological studies, because the two lines in each pair showed similar growth, N assimilation, and flowering traits under nonsymbiotic conditions. Tests of those lines under N₂-dependent conditions with isogenic Hup⁺ and negligible Hup (Hup⁻) mutants of R. leguminosarum 128C53 showed that, in symbioses with Hup⁺ rhizobia, two out of three Hup⁺⁺ pea lines decreased N₂ fixation relative to Hup⁺ peas. In one of those cases, however, the Hup⁺⁺ plant line also decreased fixation by Hup⁻ rhizobia. When results were averaged across all rhizobia tested, Hup⁺ pea isolines had 8.2% higher dry weight (P ≤ 0.05) and fixed 12.6% more N₂ (P ≤ 0.05) than Hup⁺⁺ isolines. Pea lines described here may help identify host plant factors that influence rhizobial Hup activity and should assist in clarifying how Hup systems influence other physiological processes.     

Genetic analysis and complementation studies on a number of mutant supernodulating soybean lines

Genetic analysis was done on a number of nitrate tolerant supernodulating (nts) mutant soybean lines. These lines are altered in the autoregulation response, and each was isolated as a separate mutational event following chemical mutagenesis. Crosses were made betweennts lines on a diallel pattern, and each was also crossed usingnts lines as female parent, to wild-type nodulation cultivars. F₁ and F₂ data were analysed from each cross for nodulation type and number. No complementation was noted wherents lines were intercrossed, suggesting that in each line the same gene was affected. Wherents lines were crossed with wild-type cultivars all the F₁ progeny were wild-type, confirming that thenls gene is recessive and, with one exception,nts 1116, all of the F₂ progeny segregated into a 3:1 wild-type to supernodulating phenotype, indicating that a single gene is involved. The hypernodulating linents 1116 gave a 1:1 ratio in its F₂ progeny when crossed with othernts lines. This line behaved as a dominant in the latter crosses. No wild-type segregants were recovered, therefore again no complementation look place. This line may be a leaky mutant with partial autoregulation as its segregation ratios do not fall into any of the obvious patterns.     

RT1-Linked Ir and Is genes control the immune response to bovine insulin in the rat

The immune response to bovine or pork insulin (BI or PI, respectively) was studied in the rat using the in vitro insulin-induced lymphocyte-proliferation assay. Results indicated that 11 inbred rat strains were divided into categories of high and low responders. Two high responders, SDJ (RT1 ^u) and BN(RT1 ⁿ) inbred rat strains, appeared to recognize different antigenic determinant(s) on the insulin molecule. The results of linkage and segregation analyses in F₁, F₂, backcross, and partially congenic rats showed that the Ir gene (Ir-BI), which encodes the high responsiveness in the SDJ rats, is inherited associated with RT1 ^u, whereas the immune suppression gene (Is-BI), which encodes the low responsiveness in the WKA(_RT1 ^k) rats, is inherited together with RT1 ^k. The Is-BI is the first major histocompatibility complex (MHC)-linked Is gene reported in the rat. The LEJ(RTI-A ^u B ^b) inbred rat strain showed a low response to BI, indicating that Ir-BI is closer to RTI-B/RTI-D region than to RTI-A.Abbreviations used in this paper BI bovine insulin - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - It immune response - Is immune suppression - MHC major histocompatibility complex - mol. wt. molecular weight - PI pork insulin - sc subcutaneously - SD standard deviation - SI stimulation index     

Production of alloplasmic and euplasmic wheat-barley ditelosomic substitution lines 7H<Superscript>1</Superscript>L<Superscript>mar</Superscript>(7D) and analysis of the 18S/5S mitochondrial repeat in these lines

Alloplasmic lines of common wheat with disomic substitution of chromosome 7D for telocentric chromosome 7H¹L^mar of barley H. marinum subsp. gussoneanum Hudson were isolated from the plants of generation BC₃, produced as a result of backcrossing of barley-wheat hybrids H. marinum subsp. gussoneanum (2n = 28) × T. aestivum (2n = 42), Pyrotrix, cultivar, with 28 common wheat cultivars Pyrotrix 28 and Novosibirskaya 67. Chromosome substitution pattern was determined using SSR analysis and C-banding. In preliminary genomic in situ hybridization experiments, telocentric chromosomes were assigned to wild barley was established. In the BC₃F₈ generations of three alloplasmic lines with the 7H¹L^mar(7D) substitution type the differences in fertility manifestation were observed: most of the L-32(1) plants were sterile, in line L-32(2) only sporadic plants were sterile, and line L-32(3) was fertile. Simultaneously with these experiments, using selfpollinated progeny of the hybrids obtained in crosses of common wheat cultivar Saratovskaya 29 (2n = 41), monosomic for chromosome 7D, with common wheat cultivar Pyrotrix 28 with addition of pair of telocentric chromosomes 7H¹L^mar (7D) of barley H. marinum subsp. gussoneanum, euplasmic wheat-barley ditelosomic substitution 7H¹L^mar (7D) lines were isolated. The lines obtained had normal fertility. PCR analysis of the 18S/5S mitochondrial repeat (hereafter, mtDNA sequence) in alloplasmic and euplasmic ditelosomic substitution lines 7H¹L^mar(7D) was performed. In the plants from alloplasmic sterile line L-32(1), the sequences only of the barley (maternal) type were revealed, while the plants from alloplasmic fertile lines L-32(2) and L-32(3) demonstrated heteroplasmy (the presence of barley- and wheat-like sequences within one individual). In euplasmic ditelosomic substitution lines the presence of only wheat-like 18S/5S mitochondrial repeat sequences was observed. The results indicate that the presence of barley-like mtDNA sequences in alloplasmic substitution lines was not associated with the presence of barley chromosomes in their nuclear genomes.     

Visualization of site-specific recombination catalyzed by a recombinase from Zygosaccharomyces rouxii in Arabidopsis thaliana

Excision of a DNA segment can occur in Arabidopsis thaliana by reciprocal recombination between two specific recombination sites (RSs) when the recombinase gene (R) from Zygosaccharomyces rouxii is expressed in the plant. To monitor recombination events, we generated several lines of transgenic Arabidopsis plants that carried a cryptic β-glucuronidase (GUS) reporter gene which was designed in such a way that expression of the reporter gene could be induced by R gene-mediated recombination. We also made several transgenic lines with an R gene linked to the 35S promoter of cauliflower mosaic virus. Each transgenic line carrying the cryptic reporter gene was crossed with each line carrying the R gene. Activity of GUS in F₁ and F₂ progeny was examined histochemically and recombination between two RSs was analyzed by Southern blotting and the polymerase chain reaction. In seedlings and plantlets of F₁ progeny and most of the F₂ progeny, a variety of patterns of activity of GUS, including sectorial chimerism in leaves, was observed. A small percentage of F₂ individuals exhibited GUS activity in the entire plant. This pattern of expression was ascribed to germinal recombination in the F₁ generation on the basis of an analysis of DNA structure by Southern blotting. These results indicate that R gene-mediated recombination can be induced in both somatic and germ cells of A. thaliana by cross-pollination of parental transgenic lines.     

Ambient levels of UV-B in Hawaii combined with nutrient deficiency decrease photosynthesis in near-isogenic maize lines varying in leaf flavonoids: Flavonoids decrease photoinhibition in plants exposed to UV-B

Near-isogenic lines of maize varying in their genes for flavonoid biosynthesis were utilized to examine the effects of foliar flavonoids and nutrient deficiency on maximum net photosynthetic rate (P _N) and chlorophyll (Chl) fluorescence (F_v/F_m) in response to ultraviolet-B (UV-B) radiation. Plants with deficient (30 to 70 % lower N, K, Mn, Fe, and Zn) and sufficient nutrients were exposed to four irradiation regimes: (1) no UV-B with solar photosynthetically active radiation (PAR), (2) two day shift to ambient artificial UV-B, 8.2–9.5 kJ m⁻² d⁻¹ (21–25 mmol m⁻² d⁻¹); (3) continuous ambient artificial UV-B; (4) continuous solar UV-B in Hawaii 12–18 kJ m⁻² d⁻¹ (32–47 mmol m⁻² d⁻¹). The natural ratio of UVB: PAR (0.25–0.40) was maintained in the UV-B treatments. In the adequately fertilized plants, lines b and lc had higher contents of flavonoids and anthocyanins than did lines hi27 and dta. UV-B induced the accumulation of foliar flavonoids in lines hi27 and b, but not in the low flavonoid line dta or in the high flavonoid line lc. In plants grown on deficient relative to adequate nutrients, flavonoid and anthocyanin contents decreased by 30–40 and 40–50 %, respectively, and Chl a and Chl b contents decreased by 30 and 70 %, respectively. The UV-B treatments did not significantly affect P _N and F_v/F_m in plants grown on sufficient nutrients, except in the low flavonoid lines dta and hi27 in which P _N and F_v/F_m decreased by ∼15 %. P _N, F_v/F_m, and stomatal conductance decreased markedly (20–30 %) in all lines exposed to UV-B when grown on low nutrients. The decrease in F_v/F_m was 10 % less in higher flavonoid lines b and lc. The photosynthetic apparatus of maize readily tolerated ambient UV-B in the tropics when plants were adequately fertilized. In contrast, ambient UV-B combined with nutrient deficiency significantly reduced photosynthesis in this C₄ plant. Nutrient deficiency increased the susceptibility of maize to UV-B-induced photoinhibition in part by decreasing the contents of photoprotective compounds.     

Organometallic nickel(II) complexes with dithiophosphate, dithiophosphonate and monothiophosphonate ligands

The hydroxo complex [NBu₄]₂[Ni₂(C₆F₅)₄(μ-OH)₂] reacts with ammonium O,O^′-dialkyldithiophosphates, O-alkyl-p-methoxyphenyldithiophosphonate acids and ammonium O-alkylferrocenyldithiophosphonates in dichloromethane under mild conditions to give, respectively, [NBu₄][Ni(C₆F₅)₂{S(S)P(OR)₂}] (R=Me (1), Et (2), ⁱPr (3)) and [NBu₄][Ni(C₆F₅)₂{S(S)P(OR)Ar}] (Ar=p-MeOC₆H₄, R=Me (4), Et (5), ⁱPr (6); Ar=ferrocenyl; R=Me (7), Et (8), ⁱPr (9)). The monothiophosphonate nickel complexes [NBu₄][Ni(C₆F₅)₂{S(S)P(OR)(ferrocenyl)}] (R=Et (10), ⁱPr (11)) are obtained by reaction of the hydroxo complex with O-alkylferrocenyldithiophosphonate acids. Analytical (C, H, N, S), conductivity, and spectroscopic (IR, ¹H, ¹⁹F and ³¹P NMR, and FAB-MS) data were used for structural assignments. A single-crystal X-ray diffraction study of [NBu₄][Ni(C₆F₅)₂{S(S)P(OMe)(p-MeOC₆H₄)}] (4) and [NBu₄][Ni(C₆F₅)₂{S(O)P(OEt)(ferrocenyl)}] (10) shows that in both cases the coordination around the nickel atom es essentially square planar with NiC₂S₂ and NiC₂SO central cores, respectively.     

Control of the antibody response of C57BL/6 Lyt-congenic strains to the Lyt-3.1 alloantigen by a gene linked to theLyt-2 locus

Congenic anti-Lyt-3.1 sera have recently been produced by immunizing B6-Lyt-2^a mice with thymocytes from either B6-Lyt-2^a, Lyt-3^a or B6-Lyt-2^a, Lyt-3^a, H-2^k mice (Boos et al. 1978). Surprisingly, mice of the congenic strain B6 failed to produce either anti-Lyt-2.1 or anti-Lyt-3.1 cytotoxic antibodies after identical immunizations. To determine the genetic basis for the difference in response to Lyt-3.1, (B6 × B6-Lyt-2^a)F_a mice and progeny of the backcross, (B6 × B6-Lyt-2^a)F₁ × B6-Lyt-2^a, were immunized with B6-Lyt-2^a, Lyt-3^a, H-2^k thymocytes. In addition, thymic biopsies of backcross progeny were performed and thymocytes tested for the Lyt-2.2 antigenic specificity. Results indicate that gene(s) governing the immune response to Lyt-3.1 is (are) linked to theLyt-2 locus, and that the responder allele (linked toLyt-2 ^a ) shows very poor penetrance in Lyt-2^a/Lyt-2^b mice.     

Experimentally calibrated computational chemistry of tryptophan hydroxylase: Trans influence, hydrogen-bonding, and 18-electron rule govern O2-activation

Insight into the nature of oxygen activation in tryptophan hydroxylase has been obtained from density functional computations. Conformations of O₂-bound intermediates have been studied with oxygen trans to glutamate and histidine, respectively. An O₂-adduct with O₂trans to histidine (O_his) and a peroxo intermediate with peroxide trans to glutamate (P_glu) were found to be consistent (0.57-0.59 mm/s) with experimental Mössbauer isomer shifts (0.55 mm/s) and had low computed free energies. The weaker trans influence of histidine is shown to give rise to a bent O₂ coordination mode with O₂ pointing towards the cofactor and a more activated O-O bond (1.33 Å) than in O_glu (1.30 Å). It is shown that the cofactor can hydrogen bond to O₂ and activate the O-O bond further (from 1.33 to 1.38 Å). The O_his intermediate leads to a ferryl intermediate (F_his) with an isomer shift of 0.34 mm/s, also consistent with the experimental value (0.25 mm/s) which we propose as the structure of the hydroxylating intermediate, with the tryptophan substrate well located for further reaction 3.5 Å from the ferryl group. Based on the optimized transition states, the activation barriers for the two paths (glu and his) are similar, so a two-state scenario involving O_his and P_glu is possible. A structure of the activated deoxy state which is high-spin implies that the valence electron count has been lowered from 18 to 16 (glutamate becomes bidentate), giving a “green light” that invites O₂-binding. Our mechanism of oxygen activation in tryptophan hydroxylase does not require inversion of spin, which may be an important observation.     

In vitro correction of antigen-induced immune suppression: effects of poly(A) poly(U) and prostaglandin E

Incubation of SJL or DBA/1 mouse spleen cells with poly(lTyr, lGlu)-polylPro—polylLys, (T, G)-Pro—L in vitro reduced the immune response potential of the cells to this immunogen as tested by adoptive transfer into irradiated, syngeneic recipients, followed by immunization with (T, G)-Pro—L in complete Freund's adjuvant. This reduction in immunocompetence was antigen-specific, since incubation with another antigen (rabbit immunoglobulin G) did not result in a suppression of responsiveness of the cells to subsequent in vivo immunization with (T, G)-Pro—L. Incubation of the spleen cell-(T, G)-Pro—L mixture in the presence of either prostaglandin E₁(PGE₁) or polyadenylic-polyuridylic acid (poly(A)·poly (U)) restored the immune response potential to the normal level. Incubation of (T, G)-Pro—L with spleen cells had no effect on cyclic AMP accumulation, whereas incubation of PGE₁ with the cells stimulated cyclic AMP production, irrespective of the presence of antigens. In contrast, the level of cyclic AMP was not affected by poly(A) · poly(U). The difference in cyclic AMP accumulation suggests that PGE₁ and poly(A) · poly(A) modify immune responsiveness by different mechanisms. The above observations were verified both in SJL and DBA/1 mice, which are the respective genetic high and low responders to (T, G) -Pro—L. This implies that the modifications of responsiveness described are not related to the genetic control of immune response to this immunogen.     

Overexpression of Iron Superoxide Dismutase in Transformed Poplar Modifies the Regulation of Photosynthesis at Low CO2 Partial Pressures or Following Exposure to the Prooxidant Herbicide Methyl Viologen

Chloroplast-targeted overexpression of an Fe superoxide dismutase (SOD) from Arabidopsis thaliana resulted in substantially increased foliar SOD activities. Ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase activities were similar in the leaves from all of the lines, but dehydroascorbate reductase activity was increased in the leaves of the FeSOD transformants relative to untransformed controls. Foliar H₂O₂, ascorbate, and glutathione contents were comparable in all lines of plants. Irradiance-dependent changes in net CO₂ assimilation and chlorophyll a fluorescence quenching parameters were similar in all lines both in air (21% O₂) and at low (1%) O₂. CO₂-response curves for photosynthesis showed similar net CO₂-exchange characteristics in all lines. In contrast, values of photochemical quenching declined in leaves from untransformed controls at intercellular CO₂ (Ci) values below 200 μL L⁻¹ but remained constant with decreasing Ci in leaves of FeSOD transformants. When the O₂ concentration was decreased from 21 to 1%, the effect of FeSOD overexpression on photochemical quenching at limiting Ci was abolished. At high light (1000 μmol m⁻² s⁻¹) a progressive decrease in the ratio of variable (F_v) to maximal (F_m) fluorescence was observed with decreasing temperature. At 6^oC the high-light-induced decrease in the F_v/F_m ratio was partially prevented by low O₂ but values were comparable in all lines. Methyl viologen caused decreased F_v/F_m ratios, but this was less marked in the FeSOD transformants than in the untransformed controls. These observations suggest that the rate of superoxide dismutation limits flux through the Mehler-peroxidase cycle in certain conditions.