Cardiac allografts in mice congenic at non-H-2 histocompatibility loci

Neonatal mouse heart fragments were grafted under the ear skin of adult recipients. Cardiac allograft survival was evaluated by visual observation of pulsation, electrocardiography, and histology. Employing a series of congenic resistant strains differing from C57BL/10Sn at theH-1, H-3,H-4, H-7, H-8, H-9, H-10, H-11, andH-12 loci, the median survival times of the heart grafts to and from C57BL/10Sn were obtained. The various interallelic combinations resulted in a wide variation of graft survival. Reciprocal transplants frequently showed different survival times.H-1 ^c grafts were rejected by B10.129(5M)/nSn female mice with a median survival time of 90 days.H-1 ^b grafts were not rejected by C57BL/10Sn mice for the experiment's duration of 200 days. The weaker the histocompatibility barrier, the more variable the survival times and the smaller the ratio of rejected to total grafted heart fragments. Female recipients were observed to reject their grafts more rapidly and to reject a higher proportion than males of the same strain. Although the strength of the different non-H-2 barriers generally paralleled that determined by skin transplants, the rankings of the strongest minor barriers were not the same for both tissues.     

Development of Transplantation Immunity and Restoration Experiments in the Thymectomized Amphibian

This paper examines the thymic dependence of alloimmunity inamphibians. In Xenopus, the presence of a thymus during thefirst 2 weeks of life is essential for the development of normalfirst-set skin allograft immunity. Thymectomy during this earlyperiod always impairs the alloimmune response of young adulttoads. However, most of these thymectomized animals are ableto completely destroy skin allografts, albeit with prolongedrejection times. Chronic graft rejection, rather than tolerance,still occurs following thymectomy as early as 5 days, when thethymus contains no small lymphocytes. In contrast to the considerabledifferences in first-set allograft survival times in controland early-thymectomized Xenopus, second-set grafts, appliedsubsequent to first-set destruction, are rejected in acute fashion(<3 weeks) in both groups. That the defect in first-set alloimmunityis specifically related to absence of thymus has been confirmedby implanting allogeneic thymus 2 weeks post-thymectomy. Thedonor thymus remains healthy and restores the allograft responseto normal. In contrast, allogeneic spleen does not reconstituteand itself often undergoes destruction. Preliminary autoradiographicexperiments on lymphoid tissue involvement in first-set allograftrejection are also described.     

Production of θ Antibody by Mice that Fail to Reject θ-Incompatible Skin Grafts

EVIDENCE that female mice failing to reject male skin grafts are nevertheless producing H-Y antibody indicates that graft survival in this case is not due to failure of the recipient to respond to graft antigens¹. Survival of skin grafts in recipients that are producing specific anti-graft antibody is known in another system: radiation chimaeras of constitution (C57BL × A)→A often fail to reject C57BL skin grafts² although they are producing Sk alloantibody³. Sk alloantigens are limited to epidermal cells and brain and so give rise to skin-selective homograft reactions by radiation chimaeras^2,3.     

Antigraft responses to the H-28c antigen by B6 and B6D2F1 mice

The survival of minor H antigen-bearing skin grafts from donors congenic with C57BL/6 (B6) was compared in B6, B6D2, and AB6 hybrid recipients. In a case singled out for further study, B6 mice were found to reject HW 110 skin (H-28^c antigen) rapidly, whereas B6D2 mice rejected HW110 skin much more slowly and variably. Both major histocompatibility complex (MHC)-linked and non-MHC genes appeared to affect the survival of HW110 strain skin grafts on B6 and B6D2 recipients. Results of several experiments appear to rule out the sharing of H-28° epitopes between donors and recipients as an explanation for the relatively poor response of B6D2 mice to HW 110 skin grafts. Experiments involving bone marrow chimeras produced by the reciprocal exchange of bone marrow between irradiated B6 and B6D2 mice suggest that bone marrow-derived donor cells and non-bone-marrow-derived host cells each contribute to the immune response phenotype with respect to the H-28° antigen. An attempt was made to determine whether B6D2 mice that failed to reject HW110 strain skin grafts possessed suppressor cells specific for the H-28^c antigen. Spleen cells from poorly responsive B6D2 mice failed to suppress the rejection of HW 110 skin grafts when assayed in immunodeficient mice that were provided with cells from immune 136132 donors that were highly responsive to HW110 skin grafts.     

Strong and weak immune responses across the same major histocompatibility barrier in rats

Inbred rat strains, Fischer 344 (F-344) and Lewis (LEW), share the serologicalAg-Bl allele and react very weakly in mixed lymphocyte culture (MLC). Despite this apparent identity atAg-B, these strains differ markedly in their immune responses to anAg-B disparate third strain Marshall 520 (M-520) (Ag-B6). F-344 recipients allowed M-520 heart grafts an extended survival, whereas LEW recipients rejected them rapidly. F-344 and M-520 showed a weak response in MLC in contrast to a strong response for LEW and M-520. F-344 produced antisera in response to injection of M-520 cells that had a relatively high antibody titer but low cytotoxic activity. F-344 responded to another strain, Buffalo (BUF) (alsoAg-B6), in a similar fashion. F-344 apparently can produce a strong allogeneic response, as it was able to rapidly reject heart grafts from (LEW x Brown-Norway) F₁ donors (LBN) (Ag-B 1/3). The low response of F-344 to M-520 probably was not due to shared antigens between the two strains because M-520 heart grafts underwent rapid rejection in LEW hosts highly tolerant to F-344. To explain the contrasting response of F-344 and LEW to theAg-B6 disparity, we propose that it is controlled by an immune-response gene(s); that F-344 has a low-responding allele and LEW has a high-responding allele. The data do not reveal a location for this proposed gene. The high-responding allele appears to be dominant, as M-520 hearts were rejected rapidly by (F-344 x LEW) F₁ recipients.     

Production of H-Y Antibody by Female Mice that Fail to Reject Male Skin

WHEN inbred mice are grafted with skin from inbred donors that differ from the recipients only by a single minor histocompatibility antigen, it is commonly observed that some recipients will retain their skin grafts while others will reject them. This is true of incompatibility for H-Y antigen, which is responsible for the rejection of male grafts by otherwise histocompatible inbred females of the same inbred strain¹. Thus in the DBA/2 (DBA) strain, male-to-female skin grafts are rejected by only some recipients; in the C57BL (B6) strain, females always reject male skin; and C3H/An (C3H) females usually accept male skin grafts indefinitely.     

Microvascular Regulation of Cutaneous Gas Exchange in Amphibians

SYNOPSIS. Gas exchange across amphibian skin is regulated bythe cutaneous microcirculation. Parameters involved in regulatinggas exchange are capillary density, radius and blood flow. Changesin capillary density and radius should affect gas exchange byaltering the cutaneous diffusing capacity (D₂) while changesin capillary blood flow affect the perfusive conductance ofthe skin. A simple model predicts that the effect of capillary densitychanges on D₂ will become more pronounced as capillary densityand epidermal thickness decrease. Changes in capillary radiusshould have only a minor effect on D₂ Previous analyses havesuggested that cutaneous gas exchange is not significantly affectedby the perfusive conductance of the skin. Consequently, it hasbeen thought that changes in total capillary blood flow havelittle impact on cutaneous gas exchange. Earlier analyses, however,may have underestimated the importance of perfusive conductancein amphibian skin, primarily because functional heterogeneitiesin the microcirculation were not considered. The density of perfused capillaries is regulated in the footweb of Rana esculenta by environmental Po₂ and PCO₂, and alsoby lung ventilation. In Rana catesbeiana, capillary densityin the web decreases during air exposure. Chronic exposure toenvironmental hypoxia increases total capillary density in bullfrogtadpole skin. In Rana pipiens, regulation of cutaneous gas exchangeby environmental and pulmonary O₂ probably involves changesin total capillary blood flow.     

The influence of graft size on the induction of immunity versus tolerance to H-Y in H-2 k strains of mice

Unprimed female CBA mice do not reject large (10 mm²) syngeneic male skin grafts. However, a high proportion do reject small (4 mm²) grafts. Nevertheless, rejection does not invariably result in an anamnestic response. In some cases, the immunity induced by the rejection of a small graft was overcome, and tolerance was induced by a subsequent challenge with a large graft. This suggests that the transplantation response to minor antigens is subject to active regulation, and screening of other H-2 ^k strains indicates that the nature of the response (i. e., immunity or tolerance) is determined by a gene or genes mapping outside the major histocompatibility complex.     

Antimicrobial Peptide defenses in amphibian skin

One of the most urgent problems in conservation biology todayis the continuing loss of amphibian populations on a globalscale. Recent amphibian population declines in Australia, CentralAmerica, the western United States, Europe, and Africa havebeen linked to a pathogenic chytrid fungus, Batrachochytriumdendrobatidis, which infects the skin. The skin of amphibiansis critical for fluid balance, respiration, and transport ofessential ions; and the immune defense of the skin must be integratedwith these physiological responses. One of the natural defensesof the skin is production of antimicrobial peptides in granularglands. Discharge of the granular glands is initiated by stimulationof sympathetic nerves. To determine whether antimicrobial skinpeptides play a role in protection from invasive pathogens,purified antimicrobial peptides and natural peptide mixturesrecovered from the skin secretions of a number of species havebeen assayed for growth inhibition of the chytrid fungus. Thegeneral findings are that most species tested have one or moreantimicrobial peptides with potent activity against the chytridfungus, and natural mixtures of peptides are also effectiveinhibitors of chytrid growth. This supports the hypothesis thatantimicrobial peptides produced in the skin are an importantdefense against skin pathogens and may affect survival of populations.We also report on initial studies of peptide depletion usingnorepinephrine and the kinetics of peptide recovery followinginduction. Approximately 80 nmoles/g of norepinephrine is requiredto deplete peptides, and peptide stores are not fully recoveredat three weeks following this treatment. Because many specieshave defensive peptides and yet suffer chytrid-associated populationdeclines, it is likely that other factors (temperature, conditionsof hydration, "stress," or pesticides) may alter normal defensesand allow for uncontrolled infection.     

Xanthine derivatives without PDE effect stimulate voltage-activated chloride conductance of toad skin

The effect of xanthine derivativeson the voltage-activated Cl conductance(G_Cl) of amphibian skin was analyzed.3-Isobutyl-1-methylxanthine (IBMX) and the recently synthesizedxanthine derivatives 3,7-dimethyl-1-propyl xanthine (X-32) and3,7-dimethyl-1-isobutyl xanthine (X-33), which lack inhibitory effectson phosphodiesterases in CHO and Calu-3 cells, increasedvoltage-activated G_Cl without effect on baseline conductance at inactivating voltage. Half-maximal stimulation ofG_Cl occurred at 108 ± 9 µM for X-32 andX-33 after apical or basolateral application. The stimulation ofG_Cl, which occurs only in the presence ofCl in the mucosal solution, is caused by a shift of thevoltage sensitivity to lower clamp potentials and an increase of themaximally activated level. Furosemide reversed both the shift ofsensitivity and the increase in magnitude. These patterns arefundamentally different from those seen after application ofmembrane-permeant, nonmetabolized analogs of cAMP, and they indicatethat the xanthines stimulate G_Cl directly. Thisnotion is strengthened by the lack of influence on intracellular cAMPcontent, which is consistent with the observations in CHO and Calu-3cells. We propose that the xanthine derivatives increase the voltagesensitivity of a regulative component in the conductiveCl pathway across amphibian skin.

     

The differentiation of bone marrow cells to functional T lymphocytes following implantation of thymus grafts and thymic stroma in nude and AT×BM mice

Cardiac allografts were used to compare the immunologic capacity of nude mice and adult, thymectomized, lethally irradiated, bone marrow-reconstituted (AT × BM) mice. Neither nude nor AT × BM mice were able to reject cardiac allografts of any party. However, both rejected grafts of any party following implantation of neonatal thymus or thymus from 3-week-old syngeneic mice. Irradiated syngeneic thymus grafts (800 R) were equally effective in restoring host responsiveness against allografts. In contrast, allogeneic thymus grafts restored the capacity to reject second-party heart grafts only in AT × BM mice. Second-party grafts persisted indefinitely when placed on nude mice implanted with an allogeneic, unirradiated thymus graft. Third-party grafts transplanted 17 weeks after reconstitution, however, were rejected. Irradiated nude mice given normal littermate bone marrow and simultaneously grafted with second-party thymus and heart allografts also failed to reject their second-party heart grafts. The difference in ultimate capacity to respond between AT × BM and nude mice suggests that a maturational defect exists in the nude mouse enviroment which impedes development of precursor T lymphocytes.     

Effects of Tail Clipping on Larval Performance and Tail Regeneration Rates in the Near Eastern Fire Salamander,Salamandra infraimmaculata

Tail-tip clipping is a common technique for collecting tissue samples from amphibian larvae and adults. Surprisingly, studies of this invasive sampling procedure or of natural tail clipping – i.e., bites inflicted by predators including conspecifics - on the performance and fitness of aquatic larval stages of urodeles are scarce. We conducted two studies in which we assessed the effects of posterior tail clipping (~30 percent of tail) on Near Eastern fire salamander (Salamandra infraimmaculata) larvae. In a laboratory study, we checked regeneration rates of posterior tail-tip clipping at different ages. Regeneration rates were hump-shaped, peaking at the age of ~30 days and then decreasing. This variation in tail regeneration rates suggests tradeoffs in resource allocation between regeneration and somatic growth during early and advanced development. In an outdoor artificial pond experiment, under constant larval densities, we assessed how tail clipping of newborn larvae affects survival to, time to, and size at metamorphosis. Repeated measures ANOVA on mean larval survival per pond revealed no effect of tail clipping. Tail clipping had correspondingly no effect on larval growth and development expressed in size (mass and snout-vent length) at, and time to, metamorphosis. We conclude that despite the given variation in tail regeneration rates throughout larval ontogeny, clipping of 30% percent of the posterior tail area seems to have no adverse effects on larval fitness and survival. We suggest that future use of this imperative tool for the study of amphibian should take into account larval developmental stage during the time of application and not just the relative size of the clipped tail sample.     

Locomotion in Amphibian Larvae (or "Why Aren't Tadpoles Built Like Fishes?")

SYNOPSIS. A variety of morphological features that affect locomotiondistinguish larvae of the three living amphibian orders fromfishes and their larvae. The oddest amphibian larvae are anurantadpoles. With their globose bodies, concealed forelimbs, abruptlycompressed and terminally tapered tails, tadpoles not only differradically from fishes but they—unlike caecilians or salamanders—alsodiffer radically from their adults. Tadpoles typically haveless axial musculature and much simpler myotomes than fishes.Surprisingly, in terms of mechanical (propeller) efficiencyand maximum sprint speeds, tadpoles still perform as well asmany teleosts of comparable sizes. From a consideration of hydromechanics,no amphibian larvae appear to be designed for sustained swimmingat high speeds. High maneuverability, rather than sustainablespeed, are important for amphibian larval survival.Two key featuresof tadpoles are the absence of caudal vertebrae and unexposedpectoral appendages. With only a notochord to serve as a skeleton,the tadpole tail is extremely flexible. Because of this exceptionalflexibility, tadpoles can fold their tails up against the bodyand turn rapidly with virtually no displacement of their centerof mass. Caudal flexibility can be regulated by muscle activityin the tadpole to effect turning. Lateral appendages are notneeded for this movement and are free to develop directly intotheir adult morphology; the anterior ones develop under coverof an opercular fold where they do not contribute to drag. Acase is presented, based on the ecology of metamorphosis, thatanuran transformation should be as brief as possible. With nobone to resorb, metamorphosis of the anuran caudal appendagecan, indeed, be very rapid.The basic kinematics of constantvelocity straightforward swimming for tadpoles and salamanderlarvae is reviewed, as well as the kinematics and electromyographyof starting, stopping, and turning in tadpoles. An attempt ismade to relate swimming kinematics to the characteristic morphologiesof amphibian larvae. Swimming speed in Rana, Bufo and Aynbystomalarvae, which swim only intermittently, is modulated by changingtail beat frequency. However, Xenopus, which swims constantlyby sculling with its tail, regulates swimming speed (at lowto intermediate velocities) by varying the length of the propulsivewave in its tail. Xenopus and Rana differ in the morphologyof their notochord, spinal cord, spinal nerves, and spinal motorpool distribution within the spinal cord. These differencesmay underlie the different way these larvae regulate swimming.They may also reflect their phylogenetic history.     

Characterization of a wilty sunflower (Helianthus annuus L.) mutant: III. Phenotypic interaction in reciprocal grafts from wilty mutant and wild-type plants

The present study was conducted to evaluate phenotypic interactionin reciprocal grafts between wilty (w-1) sunflower mutant andnormal (W-1) plants. The w-1 genotype is a ‘leaky’ABA-deficient mutant, characterized by high stomatal conductance,in both light and dark conditions, and high transpiration rate. In well-watered conditions, mutant scions grafted on to normalrootstock (w-1/W-1) showed higher leaf relative water content,leaf water potential and ABA levels than those of control grafts(w-1/w-1). In addition, detached leaves of w-1/W-1 exhibitedlower water loss than w-1/w-1 grafts, while mutant rootstockdid not affect the transpiration rate of detached W-1 leaves.When drought stress was imposed to potted plants by withholdingwater, the mutant scions grafted on to normal roots showed apartial phenotypic reversion. A rapid stomatal closure and arise in ABA levels in response to a small decrease in leaf waterpotential was observed. By contrast, in w-1/w-1 grafts significantreductions in stomatal conductance and ABA accumulation weredetected only in conjunction with a severe water deficit. W-1scions on mutant stocks (W-1/w-1) maintained the normal phenotypeof control wild-type grafts (W1/W-1). Key words: ABA, grafting, Helianthus annuus, stomatal conductance, water relations, wilty mutant     

Transplantation Immunity in Annelids. III. Effects of Temperature on Xenograft Rejection in Earthworms

Earthworms are capable of destroying body wall xenografts. Xenograftstransplanted Eisenia foetidaLumbricus terrestris at varioustemperatures showed that graft survival time is inversely proportionalto temperature in both cases. Colder temperatures slow graftrejection, but higher temperatures, up to optimal limits, accelerategraft destruction. At 21 C, Eisenia grafts on Lumbricus hostsare rejected faster than at any other experimental temperature;the variability in individual times is lessened. Beginning at23 C sub-optimal conditions are approached. Thus, graft survivaltimes are more prolonged. Lumbricus grafts on Eisenia hostsare rejected most quickly at 23 C; at 27 C all Eisenia are dead.A review of the earthworm's general thermal dependence is presentedand a rationalization for prolonged second-set graft survival(negative memory) at low temperatures is given. Temperatureeffects probably account for the lack of positive memory at15 C from previous studies.     

Evolution of Hoxa-11 Expression in Amphibians: Is the Urodele Autopodium an Innovation?

In this paper we present preliminary results on the expressionof Hoxa-11 in the frog Xenopus laevis and the newt Notophthalmusviridescens. In amniote limbs, Hoxa-11 is expressed exclusivelyin the prospective zeugopodium and is involved in the developmentof zeugopodial character identity. While the Hoxa-11 expressionpattern in Xenopus is similar to those described in mice andchick, the newt Notophthalmus has a phase of autopodial Hoxa-11expression starting with the development of digit III. Expressionis particularly strong in the buds of the postaxial digits.This Hoxa-11 expression pattern is unique among tetrapods andadds to the list of developmental peculiarities of urodele limbdevelopment, which also include an anterior to posterior polarityof digit development. We propose a scenario to explain the evolutionaryorigin of urodele limb development. We assume that recent urodelesare derived from a lineage of amphibians which underwent partialdigit reduction, leaving only two digits intact, most likelydigits III and IV according to Morse's law. This implies thaturodele digits I and II are homologous to amniote digits IIIand IV, and that the postaxial digits of urodeles are an evolutionaryinnovation (re-invention). Supporting evidence for this homologyhypothesis is found in the developmental connections betweenthe basale commune and the postaxial zeugopodial element.     

卵形鲳鲹对刺激隐核虫的免疫应答和免疫保护研究

用刺激隐核虫（Cryptocaryon irritans）的幼虫对卵形鲳鲹（Trachinotus ovatus）进行腹腔注射和体表感染,然后每隔1周用阻动试验（Immobilization assay）检测免疫鱼的抗血清和皮肤培养液对激刺隐核虫幼虫的阻动效价,在第14周,分别用亚致死剂量和致死剂量的刺激隐核虫幼虫对免疫鱼攻毒以检测所产生的免疫保护力。实验结果显示：两种免疫方法都能让卵形鲳鲹的血清和皮肤生成阻动刺激隐核虫幼虫的特异性抗体,并能使被免疫鱼获得明显的免疫保护,但是体表感染免疫组的血清和皮肤培养液的阻动效价都要比腹腔注射免疫组高,所获得的免疫保护力也更强。同时还发现,免疫鱼血清和皮肤培养液中的抗体存在明显的差异：两者的最初生成时间、达到峰值的时间、变化规律以及阻动效价等都不一致。因此,我们推测鱼类的系统免疫应答和皮肤粘膜免疫应答有可能是相互独立的,或者是不同步的。鱼类的体液免疫应答,特别是粘膜免疫应答对抵御刺激隐核虫的感染起了重要的作用,采用刺激隐核虫虫体疫苗可能成为预防海水鱼类白点病的一种选择。     

Heart, but not skin, allografts from donors lacking Flt3 ligand exhibit markedly prolonged survival time

Fms-like tyrosine kinase 3 ligand (Flt3L) administration leads to dramatic increases in dendritic cells (DC) in lymphoid and nonlymphoid tissues. Conversely, mice lacking Flt3L (Flt3L(-)/(-)) show severe reductions in both myeloid (CD11c(+)CD8alpha(-)) and lymphoid-related DC (CD11c(+)CD8alpha(+)) in the thymus and secondary lymphoid organs. In this study marked reductions in CD11c(+) interstitial cardiac DC and in dermal, but not epidermal, DC (Langerhans cells) were also observed. CD11c(+) cells that migrated from Flt3L(-/-) skin explants expressed lower surface MHC class II and costimulatory molecules and naive T cell allostimulatory activity than migratory wild-type (wt) C57BL/6 (B6) CD11c(+) cells. We examined the survival of Flt3L(-)/(-) heart or tail skin grafts (H2(b)) in allogeneic wt (BALB/c; H2(d)) recipients. The outcome of transplantation of BALB/c organs into Flt3L(-)/(-) recipients was also determined. Flt3L(-)/(-) mice rejected BALB/c heart or skin grafts with similar kinetics as B6 wt recipients. Trafficking of donor DC into host spleens or draining lymph nodes was markedly reduced after transplantation of Flt3L(-)/(-) heart, but not skin grafts, respectively. Compared with wt hearts, survival of Flt3L(-)/(-) hearts was markedly prolonged in BALB/c recipients (median survival time, 37 and 15 days, respectively; p < 0.001). Skin graft survival was unaffected. Rejection of Flt3L(-/-) hearts was precipitated by infusion of wt donor DC at the time of transplant. Thus, severe depletion of interstitial heart DC resulting from targeted gene disruption prolongs, but does not indefinitely extend, heart survival. Acute rejection of wt grafts in Flt3L(-/-) recipients reflects presumably an intact role of the direct pathway of allorecognition.     

水蛭捕食两栖类卵

许多卵生动物具有对付食卵者的精巧策略以降低死亡率,因为早期发育阶段的捕食压力能显著影响具复杂生活史动物的生存。两栖动物卵被水蛭等很多物种所捕食。我们查阅了水蛭捕食两栖类卵的文献,发现27篇论文。在所报道的水蛭捕食两栖类卵的例子中,无尾类(3.6%,n=591)明显多于有尾类(1.6%,n=255)。此外,我们第一次记录到水蛭捕食四趾螈(Salamandrina perspicillata)卵,这也是西欧有尾类的第一例报道。我们没有发现两个两栖动物类群间存在差异。然而,当我们分别分析来自北美、欧洲和亚洲三个地区的数据时,发现北美的有尾类和无尾类间存在显著差异。水蛭可捕食各生活史阶段的两栖动物,因此,水蛭应归为一些两栖类的天敌。