Genetic basis of the effects of ultraviolet light B on cutaneous immunity. Evidence that polymorphism at the Tnfa and Lps loci governs susceptibility

The ability of local ultraviolet B (UVB) irradiation to impair the induction of dinitrofluorobenzene (DNFB)-specific contact hypersensitivity (CH) in mice has been shown to be genetically determined. We have explored the possibility that the mouse Tnfa and Lps loci are involved. We demonstrate that C3H/HeN (Lps ⁿ strains are UVB-susceptible, whereas C3H/HeJ (Lps ^d ) strains are UVB-resistant. Our results indicate that local intradermal (ID) injection of mouse recombinant tumor necrosis factor-alpha (TNF_a) into sites painted with DNFB impaired the induction of CH, and in a dose response experiment the effect was found to be more marked in C3H/HeN than in C3H/HeJ. Systemic administration of neutralizing TNF_a-specific antibody reconstituted the UVB-induced defect in induction of CH in UVB-susceptible mice, confirming that TNF_a is a major mediator of the deleterious effects of UVB on induction of cutaneous immunity. The UVB-susceptibility trait (revealed by effects on CH) correlates positively with a recently described restriction fragment length polymorphism (RFLP) at the Tnfa locus (allele b) and with the wild-type Lps ⁿ allele. These results suggest that appropriate alleles at the Tnfa and Lps loci conspire to render mice susceptible to the impairment of CH induction by UVB. We propose that the mechanism may function through the capacity of UVB to elicit excessive local (cutaneous) production of TNF_a, which mediates the immune defect.     

cis-urocanic acid suppression of contact hypersensitivity induction is mediated via tumor necrosis factor-alpha.

Ultraviolet B (UVB) light impairs the induction of contact hypersensitivity to epicutaneously applied haptens in certain strains of mice by a genetically determined mechanism that depends upon the participation of TNF-alpha. Because the superficial epidermis contains large amounts of trans-urocanic acid (trans-UCA), because exposure to UVB radiation converts this compound to cis-UCA, and because cis-UCA has been reported to be immunosuppressive, we have examined the possibility that the TNF-alpha-dependent effects of UVB on contact hypersensitivity induction in mice are mediated via conversion of trans- to cis-UCA. By injecting cis-UCA intradermally before application of dinitrofluorobenzene, by treating cis-UCA-injected mice systemically with neutralizing anti-TNF-alpha antibodies, and by comparing the consequences of these maneuvers in UVB-susceptible and UVB-resistant strains of mice, we have determined a) that cis-UCA can impair the induction of contact hypersensitivity in a manner similar to UVB radiation, and that the impairment is dependent upon TNF-alpha; b) that cis-UCA altered the morphology of epidermal Langerhans cells in a manner similar to UVB radiation, and that the alteration was dependent, in part, upon TNF-alpha; and c) that the inhibitory effects of cis-UCA on induction of contact hypersensitivity and the histologic effects of this compound on epidermal Langerhans cells appear to be influenced by alleles at the Tnf alpha and Lps loci. Based on these findings we propose that UVB radiation impairs the induction of contact hypersensitivity in mice by converting trans-urocanic acid to cis-UCA within the epidermis; cis-UCA in turn causes the local release of TNF-alpha, which thwarts sensitization by its ability to trap epidermal Langerhans cells transiently within the epidermis, and thereby prevents the immunogenic signal from reaching the draining lymph node where activation of unprimed, Ag-specific T cells must occur.     

Polymorphisms in the TNF region confer susceptibility to UVB-induced impairment of contact hypersensitivity induction in mice and humans

Acute, low-dose ultraviolet B radiation protocols impair induction of contact hypersensitivity (CHS) to highly reactive haptens in some mice and humans (UVB-susceptible) but not others (UVB-resistant). These deleterious effects of ultraviolet radiation appear to be mediated in part by tumour necrosis factor alpha (TNF-alpha), which is released from, or accumulates in, UVB-exposed skin. To test the hypothesis that a polymorphism of the Tnfa locus governs the UVB-S and UVB-R phenotypes, studies have been conducted in genetically disparate strains of mice. Mice carrying the Tnf(d) allele [with precisely 14 (CA) repeats in the promoter region] display the UVB-R phenotype, whereas mice with different Tnf alleles [with (CA) repeats of 14] display the UVB-S phenotype. Molecular genetic studies of the TNF region of HLA in humans displaying either the UVB-S or UVB-R phenotype reveal a significant increase in the frequencies of TNFa2 in UVB-S individuals (P=0.00032) and of TNFd3 in UVB-R individuals (P=0.012). Moreover, DNA sequencing analyses of five single-nucleotide polymorphisms (SNPs) of the TNF promoter region revealed a significant increase in the frequency of TNF/-863A (P=0.015). We propose that the TNF region dictates susceptibility to the deleterious effects of UVB radiation on the induction of contact hypersensitivity in both mice and humans, and that the UVB-S-promoting polymorphisms significantly promote the risk of sunlight-induced skin cancer in humans.     

Suppression of the cutaneous immune response following topical application of the prostaglandin PGE2

UVB irradiation (290-320 nm) and topical applications of arachidonic acid (AA) in mice decrease the number of identifiable Langerhans cells and alter the cutaneous immune response. Application of contact allergens such as dinitrofluorobenzene (DNFB) to irradiated or AA-treated skin induces antigen-specific tolerance. Indomethacin (IM), a cyclooxygenase inhibitor, administered orally to mice prior to UVB irradiation or prior to the topical application of arachidonic acid, abrogates suppression of contact hypersensitivity (CHS) to DNFB. This suggests a byproduct of arachidonic acid generated through the cyclooxygenase pathway may be involved in the immune suppression. Topical application of various prostaglandins (PGE2, PGD2, PGF2 alpha, and CTXA2) did not cause alterations in the population density of the identifiable Ia+ dendritic Langerhans cells. PGE2, but no other tested agent, produced a suppression of the CHS response to DNFB. These observations suggests that of the various prostaglandins, PGE2 might be one of several biochemical signals which mediate the suppression of contact hypersensitivity reactions following ultraviolet radiation exposure. However, the mechanisms by which PGE2 produces its suppressive effects have not been identified.     

Regulation of ultraviolet radiation induced cutaneous photoimmunosuppression by Toll-like receptor-4

UVB radiation is a potent immunosuppressive agent that inhibits cell-mediated immune responses. The mechanisms by which UVB radiation influences cell-mediated immune responses have been the subject of extensive investigation. However, the role of innate immunity on photoimmunological processes has received little attention. The purpose of this study was to determine whether Toll-like receptor-4 (TLR4) contributed to UV-induced suppression of contact hypersensitivity (CHS) responses. TLR4^−/− and wild type C57BL/6 (TLR4^+/+) mice were subjected to a local UVB immunosuppression regimen consisting of 100 mJ/cm² UVB radiation followed by sensitization with the hapten DNFB. Wild type TLR4^+/+ mice exhibited significant suppression of contact hypersensitivity response, whereas TLR4^−/− developed significantly less suppression. The suppression in wild type TLR4^+/+ mice could be adoptively transferred to naïve syngeneic recipients. Moreover, there were significantly fewer Foxp3 expressing CD4⁺CD25⁺ regulatory T-cells in the draining lymph nodes of UV-irradiated TLR4^−/− mice than TLR4^+/+ mice. When cytokine levels were compared in these two strains after UVB exposure, T-cells from TLR4^+/+ mice produced higher levels of IL-10 and TGF-β and lower levels of IFN-γ and IL-17. Strategies to inhibit TLR4 may allow us to develop immunopreventive and immunotherapeutic approaches for management of UVB induced cutaneous immunosuppression.     

Regulation by the H-2 gene complex of delayed type hypersensitivity

Identity at the major histocompatibility complex (MHC) was essential for successful transfer of delayed type hypersensitivity (DTH) in mice. The regions of the MHC involved differed according to the antigen used for sensitization. In the case of fowl gamma globulin (FGG), identity atI-A was necessary, whereas with dinitrofluorobenzene (DNFB), identity at theK, I, orD region was sufficient. These different genetic constraints probably reflect differences in the mechanisms by which antigens are presented to T lymphocytes. Cells from sensitized (CBA×C57BL)F₁ mice transferred DTH to FGG into parental-strain mice, but transfer was more effective in C57BL than in CBA with the same cell dose. This phenomenon is governed by the MHC, since there was better transfer intoH-2 ^b than intoH-2 ^k mice, regardless of their backgrounds. It may reflect the activity of an Ir gene-dependent process. Cells of one genotype (e.g., CBA), sensitized in chimeric mice derived from two MHC-incompatible strains (CBAC57BL), transferred DTH to both strains. These results do not support the notion that the genetic constraint observed in DTH transfer may be a result of the necessity for sensitized T and stimulator cells to match an identical MHC-coded cell interaction molecule. Rather, they favor the hypothesis that T cells recognize antigen, not as a naked determinant, but in close association with products of genes of the MHC.     

Low dose ultraviolet B-irradiated Langerhans cells preferentially activate CD4+ cells of the T helper 2 subset

UVB radiation distorts the Ag-presenting function of epidermal Langerhans cells (LC); this has been shown for the presentation of soluble Ag to primed T cells in vitro and for the initiation of delayed-type hypersensitivity in vivo, such as contact hypersensitivity (CH). Previous work has also demonstrated UVB-induced suppression of CH to be mediated ultimately by T cells. Two subsets of CD4+ Th cells, Th1 and Th2, have been identified, based on their cytokine production and functional activities. In particular, Th1 mediate delayed-type hypersensitivity, whereas Th2 do not. To investigate whether the perturbation of LC function induced by UVB radiation leads to a differential activation of these subsets of CD4+ cells, we examined the capacity of unirradiated and irradiated (200 J/m2) APC from adult BALB/c mice to present keyhole limpet hemocyanin to Ag-specific, H2d-restricted Th1 and Th2 cell lines. Four sources of APC were utilized: epidermal cells (EC), flow microfluorometry-purified Ia+ EC (LC), flow microfluorometry-purified Ia- EC, and splenic adherent cells (SAC). Unirradiated EC, LC, and SAC, but not Ia-EC, presented keyhole limpet hemocyanin to both Th1 and Th2. Irradiated EC and LC lost their ability to stimulate Th1, but retained fully their capacity to stimulate Th2. On the other hand, irradiated SAC were unable to induce proliferation of either Th1 or Th2. These findings indicate that suppression of CH mediated by UVB-irradiated LC may result from an alteration of the ratio and/or activity of Th1 and Th2 cells normally generated during the induction of such responses.     

Langerhans cells that migrate to skin after intravenous infusion regulate the induction of contact hypersensitivity

Intravenous infusion of hapten-derivatized epidermal cells (EC) in syngeneic mice leads to two competing signals for contact hypersensitivity (CH), a dominant effector signal attributable to Langerhans cells (LC) and a suppressor signal from Thy-1+ EC. In vitro exposure of LC to low dose ultraviolet B (UVB) radiation before hapten-derivatization and infusion not only results in the abrogation of their effector signal but also causes the down-regulation of subsequent CH responses. To delineate the relevance of i.v. immunization to the study of CH and of LC as the immunologic targets of low dose UVB radiation, we examined the migratory and immunogenic properties of EC after i.v. infusion. Unsorted EC migrated from blood to skin and lymphoid tissues, reaching steady state distributions at 16 h after infusion. No significant differences were observed between the trafficking of EC in syngeneic and allogeneic transfers. LC localized preferentially to skin, whereas Thy-1+ EC trafficked to skin, the thymus, mesenteric lymph nodes, and spleen. The pattern of trafficking of unirradiated and low dose UVB-irradiated LC were identical, suggesting that low dose UVB radiation had little effect on LC migration. Finally, skin graft experiments demonstrated i.v. infused, hapten-derivatized LC that migrate to skin to retain their capacity to induce CH, a property that was converted by in vitro pretreatment with low dose UVB radiation into down-regulation. These findings confirm the relevance and utility of the i.v. immunization model in the study of CH and the influence of low dose UVB on this immune response. Our data also provide a basis for investigating the role of disparate trafficking patterns in generating effector and suppressor signals when hapten-derivatized EC are employed for CH.     

Genetic control of contact sensitivity in mice: Effect ofH-2 and nonH-2 loci

The genetic control of delayed-type hypersensitivity in mice was investigated by contact sensitization with picryl chloride. Distribution patterns of contact sensitivity in 11 inbred strains of mice showed significant differences among strains. Comparison of levels of response between congenic-resistant lines and their inbred partners, at 9 to 11 weeks of age, revealed a clear association betweenH-2 haplotype and the magnitude of response. Testing ofH-2 recombinants further suggested the influence of two genes mapping at either end of theH-2 complex. While theH-2K ^d andH-2D ^k alleles were associated with a high response, theH-2K ^k ,H-2K ^b ,H-2D ^d , andH-2D ^b alleles were associated with a low response. Analysis of the ontogeny of response suggested that theH-2 haplotype manifests its effect through the maturation of contact sensitivity. On both the C57BL/6By and C57BL/10Sn backgrounds, theH-2 ^d haplotype was associated with early maturation of response, while theH-2 ^b haplotype was associated with late maturation. Analysis of the response of congenic lines with different genetic backgrounds and of CXB recombinant-inbred lines further revealed the marked effects of yet other genes on this trait.     

Influence of contact sensitivity to DNFB on the induction of carrier-determined tolerance to DNP.

Contact sensitivity to DNFB, induced by skin painting Balb/c mice with DNFB had no influence on the induction of carrier determined tolerance to DNP. In contrast, contact sensitivity to DNFB, elicited in complete Freund's adjuvant, prevented induction of tolerance to DNP. This effect was not due to the adjuvant alone and was hapten specific since contact sensitivity induced by OCBC in complete adjuvant had no influence on tolerance induction to DNP. In addition, in mice primed with DNFB in complete Freund's adjuvant, the tolerogen becomes immunogenic. It is suggested that the T-cell mediating tolerance to DNP-autologous IgG is different from the T-cell mediating contact sensitivity to DNP autologous carrier. DNFB in complete adjuvant may augment not only contact-sensitized T-cells, which mediate contact sensitivity, but also T-cells which have helper function in the antibody response to DNP.     

A study of cells present in peripheral lymph of pigs with special reference to a type of cell resembling the langerhans cell

Summary Large mononuclear cells with long, actively moving cytoplasmic veils were observed in lymph coming from the skin. The enzyme histochemistry and ultrastructure of these cells suggested that they are related to epidermal Langerhans cells and interdigitating cells in the lymph node. It has been reported that Langerhans cells and interdigitating cells play a role in contact hypersensitivity by taking up antigen and presenting it to thymus-dependent lymphocytes, and it is likely that the veiled cells in the lymph are also involved.After skin-painting with 1-fluoro-2,4-dinitrobenzene (DNFB), the veiled cells in lymph coming from the site of painting became more active and were observed contacting other cells present in the lymph; many large cellular aggregates were found. Since neutrophilic leucocytes and mononuclear phagocytes were the predominating cell types in this lymph, there was no evidence for a massive recruitment of immunocompetent lymphocytes at the site of painting.Neonatally thymectomized pigs do not develop allergic reactivity to DNFB. It is of interest that the number of veiled cells and their ability to form large cellular aggregates was not affected in these animals. Therefore, it is unlikely that the defect in responsiveness can be attributed to a failure in the function of veiled cells.     

Delayed-type hypersensitivity to allogeneic mouse epidermal cell antigens

Footpad swelling response was used to measure the alloantigenicity of epidermal cells (ECs) in delayed-type hypersensitivity (DTH). Strong footpad swelling was oberserved 3 h after the challenge, and it continued for 48 h after the challenge. Genetical incompatibility between the recipients and the ECs was required to induce significant footpad swelling. H-2 or non-H-2 incompatibility between mice and ECs in the sensitization phase sufficed to develop significant footpad swelling. Incompatibility caused by point mutation in the A region induced strong responses when B6. C-H-2 ^bm12 mice were immunized with B6/J ECs, but the disparity in immuno-globulin h (Igh) allotype genes was insufficient. H -Y antigen on ECs could also elicit the DTH response. Semiallogeneic F₁-derived ECs sensitized the parental recipients. The responses were successfully transferred by immune lymph node cells, but not by immune sera. Treatment of these immune lymph node cells with monoclonal antibodies plus complement revealed that the cells responsible for DTH transfer were Lyt-1⁺2^–, Ia^– T cells.Abbreviations used in this paper DNFB 2,4-dinitro-1-fluorobenzene - DTH delayed-type hypersensitivity - ECs epidermal cells - HBSS Hanks' balanced salt solution - MHC major histocompatibility complex - PBS phosphate-buffered saline     

Genetic control of acquired resistance to visceral Leishmaniasis in mice

A series ofH-2 and non-H-2 congenic resistant (CR) strains on a C57BL/10Sn background were infected with 10⁷ amastigotes ofLeishmania donovani. Non-H-2 congenic strains B10.LP-H-3 ^b and B10.CE(30NX) and (B10.LP-H-3 ^b × B10)F₁ hybrids showed a very rapid decrease in liver-parasite burdens beyond day 21. Parasite counts for these strains at day 35 were significantly lower than for all other strains tested. The rapid decrease in parasite numbers, massive lymphocellular infiltration into the liver and strong delayed hypersensitivity reactions to parasite antigens in strains congenic for a portion of chromosome 2 indicated that acquired immunity toL. donovani was controlled by a dominant gene at or near theIr-2 locus. In addition, B10.129(10M) mice, which differ from C57BL/10Sn at theH-11 locus, showed highly significant increases in parasite numbers at day 35. Other observations supporting the absence of acquired immunity in B10.129(10M) included negative delayed hypersensitivity tests to parasite antigens and the absence of lymphocellular infiltrate into the liver. Although the differences were not as pronounced,H-2 CR strains withH-2 ^b ,H-2 ^a , andH-2 ^k haplotypes also showed significantly greater decreases in parasite numbers by day 35 as compared to otherH-2 CR strains.     

The induction of tolerance to DNFB contact sensitivity by using hapten-modified lymphoid cells. III. Effects of hapten concentration on the ability of MLS-disparate cells to induce rapid unresponsiveness

We investigated genetic restrictions in the induction of immediate tolerance to DNFB contact sensitivity in mice. Using spleen cells from various donor strains haptenated at 500 micro M DNFB, we were unable to detect any restrictions in tolerance induction in recipients that were either syngeneic or allogeneic to the donor strain. However, if the concentration of hapten used in the in vitro labeling was decreased (from 500 micro M to 2.5 to 5 micro M DNFB), differences in tolerogenesis between the various donor strain haplotypes were found. Haptenated spleen cells labeled with 5 micro M DNFB produced a profound level of unresponsiveness in allogeneic recipients but produced minimal tolerance in syngeneic animals. This tolerant state was shown to be antigen-specific and was not produced by unmodified allogeneic cells alone. Further genetic analysis demonstrated that an efficient tolerant state was produced when the donor of the tolerogen and recipient differed at the MLS locus rather than at either the MHC or minor regions. This phenomenon required viable, Thy 1-bearing cells in the haptenated donor population for efficient tolerogenesis to DNFB contact sensitivity.     

A genetic analysis of natural resistance to nonsyngeneic cells: The role of H-2

The genetic control of natural resistance in vivo to four natural killer (NK) cell-resistant H-2 homozygous lymphoid tumor cell lines was investigated by following the survival and organ distribution of cells prelabeled with radioactive iododeoxyuridine. Backcross mice derived from DBA/2J and CBA/J parents were injected with H-2 ^dtumor cells and tumor cell elimination was lowest in H-2 ^dhomozygotes. Natural killer cell activity was also reduced in mice with the H-2 ^dhaplotype, but no direct correlation between NK cell levels against YAC-1 or SL2-5 lymphoma cells and natural resistance in vivo was demonstrable. Analysis of 23 BXD recombinant inbred strains indicated that natural resistance to H-2 ^dtumors was restricted to H-2 ^bstrains. There was no direct association of NK cell activity with H-2 type in the BXD strains and NK cell levels did not correlate with tumor survival in vivo. By comparing natural resistance to H-2 ^dand H-2 ^btumors in DBA/2, C57BL/6, B6D2F₁, and B10.D2 mice we found that H-2 nonidentity between the tumor and the host, rather than the host H-2 haplotype, determined whether natural resistance occurred. Again, NK cell activity against YAC-1 cells was not predictive of tumor survival in these strains. These results provide genetic evidence that NK cells alone cannot account for natural resistance to H-2 nonidentical cells of hemopoietic origin.     

Control of UVB immunosuppression in the mouse by autosomal and sex-linked genes

Irradiation with UVB (290–320 nm) initiates a systemic immunosuppression detectable as suppression of contact hypersensitivity (CHS). We investigated susceptibility to UV suppression in reciprocal F₁-hybrid and backcross mice derived from BALB/c (low susceptibility) and C57BL/6 (high susceptibility) inbred strains. CB6F₁ male mice exhibited high susceptibility and B6CF₁ male mice exhibited low susceptibility, indicating a major X-linked effect in the genetic control of UV immune suppression. Females of either F₁ hybrid showed intermediate suppression, consistent with random X-inactivation. A model of monogenic X-linked control was not sufficient, and evidence for the action of two genetically unlinked autosomal genes was found in parental backcross animals. Both sexes of (BALB/c × CB6F₁) mice showed a 1 high: 1 low ratio of phenotypes, indicating control by a major autosomal locus, Uvs1, confirmed by propagation of the high phenotype through selective backcrossing for nine generations to BALB/c. Uvs1 was not genetically linked to 12 chromosomal markers including the pigment genes b (brown) and c (albino). Backcross animals (C57BL/6 × CB6F₁) showed a significant sex difference, male mice giving a 3 high: 1 low ratio of phenotypes, compatible with the action of a second autosomal locus, Uvs2, in this hybrid. The findings are compatible with a model in which high phenotype (Uvs1 ^b/Uvs1 ^b) is dominant when subjected to recessive epistatis by the X-chromosome locus Uvs3, or by the autosomal locus Uvs2. The finding of genetic control by interacting autosomal and X-linked genes is unique. Genetically determined high susceptibility to UV immunosuppression may be an important risk factor for UV-related human diseases.     

Specific suppressor T cells in rats active in the afferent phase of contact hypersensitivity

The optimal conditions for the induction of contact hypersensitivity in rats and the characteristics of its suppression were studied using the sensitizing haptens dinitrofluorobenzene (DNFB) and trinitrochlorobenzene (TNCB). The hypersensitivity was shown to be hapten specific in so far as TNCB did not sensitize for DNFB responses but sensitization with DNFB did allow a marginal response in rats challenged with TNCB. Suppression of the sensitization to DNFB and TNCB could be generated by intravenous injection of dinitrobenzenesulphonic acid (DNBS) or trinitrobenzenesulphonic acid (TNBS), respectively, up to 3 weeks before sensitization. This suppression was hapten specific and could be transferred with splenic T cells enriched for lymphocytes carrying the OX8 (Tc/s) cell marker. Only the induction phase of sensitization, however, could be suppressed in that way. No suppression acting upon the effector phase could be detected except for a nonspecific local suppression at the site of a previous challenge with an antigen to which the rat was specifically suppressed. This study shows that suppression of contact hypersensitivity in rats is mediated by specific suppressor T cells of which the activation pathway apparently differs from that postulated for mice.     

Genetic control of contract hypersensitivity. II. Suppressor cells induced by intravenous injection of DNP-coupled syngeneic spleen cells possessing thy-1 antigen, I-J determinant, Lyt-2+, 3+ antigen, FcR- and DNP-membrane binding receptor on their surfaces

The characteristics of suppressor cells induced by 2,4-dinitrophenyl (DNP)-coupled syngeneic lymphocytes (syninduced suppressor cells) were studied. 2,4-dinitro-1-fluorobenzene (DNFB) contact hypersensitivity was completely suppressed when the syninduced suppressor cells were transferred intravenously. These syninduced suppressor cells had surface markers of Thy-1, FcR^? and Lyt-2⁺, 3⁺ antigens, as well as I-J gene products on their cell surfaces. The suppression of DNFB contact hypersensitivity was abrogated when these suppressor T cells were incubated in Petri dishes coated with the DNP-syngeneic lymphoid cell membrane, which suggests that these suppressor T cells had the specific antigen-binding receptors on their cell surfaces.     

Combination treatment of elevated UVB radiation,CO2 and temperature has little effect on silver birch (Betula pendula) growth and phytochemistry

Elevations of carbon dioxide, temperature and ultraviolet‐B (UBV) radiation in the growth environment may have a high impact on the accumulation of carbon in plants, and the different factors may work in opposite directions or induce additive effects. To detect the changes in the growth and phytochemistry of silver birch (Betula pendula) seedlings, six genotypes were exposed to combinations of ambient or elevated levels of CO₂, temperature and UVB radiation in top‐closed chambers for 7 weeks. The genotypes were relatively similar in their responses, and no significant interactive effects of three‐level climate factors on the measured parameters were observed. Elevated UVB had no effect on growth, nor did it alter plant responses to CO₂ and/or temperature in combined treatments. Growth in all plant parts increased under elevated CO₂, and height and stem biomass increased under elevated temperature. Increased carbon distribution to biomass did not reduce its allocation to phytochemicals: condensed tannins, most flavonols and phenolic acids accumulated under elevated CO₂ and elevated UVB, but this effect disappeared under elevated temperature. Leaf nitrogen content decreased under elevated CO₂. We conclude that, as a result of high genetic variability in phytochemicals, B. pendula seedlings have potential to adapt to the tested environmental changes. The induction in protective flavonoids under UVB radiation together with the positive impact of elevated CO₂ and temperature mitigates possible UVB stress effects, and thus atmospheric CO₂ concentration and temperature are the climate change factors that will dictate the establishment and success of birch at higher altitudes in the future.     

Infiltration by inflammatory cells required for solar-simulated ultraviolet radiation enhancement of skin tumor growth

In this study we compared the effects of subinflammatory and inflammatory doses of solar-simulated ultraviolet (UV) radiation on enhancement of skin tumor growth, sensitization to haptens and cellular changes within the epidermis of C3H/HeN mice. Tumors transplanted into mice 3 days after exposure to inflammatory, but not subinflammatory, doses of UV radiation had a higher growth rate than those tumors inoculated into unirradiated control mice. Both doses of UV radiation suppressed the induction of contact hypersensitivity and induced tolerance when hapten was painted onto the skin 3 days after irradiation. Skin exposed to the higher, but not the lower, dose of UV radiation contained significantly increased numbers of CD11b+, CD45+ MHC class II- and CD45+ MHC class II(hi) inflammatory cells 3 days post-irradiation. The immunosuppression correlated with a reduction in Langerhans cells and dendritic epidermal T cells. Collectively, this suggests that suppression to contact sensitizers is due to the UV radiation effects on Langerhans cells and dendritic epidermal T cells. While these effects may also suppress the induction of anti-tumor immunity, at higher doses of UV radiation inflammatory cells may enhance tumor growth by a non-immunological mechanism.