Detection of Bet v1, Bet v2 and Bet v4 specific IgE antibodies in the sera of children and adult patients allergic to birch pollen: evaluation of different IgE reactivity profiles depending on age and local sensitization.

BACKGROUND: Birch pollen belongs to the major allergen triggers in the spring season in Europe. Our rapidly expanding knowledge of the allergenic molecules enables us to better recognize the individual differences between the reactivity of specific IgE antibodies of individual patients and allergic populations living in various regions of the world. METHOD: In a group of birch pollen-allergic patients living in the Czech Republic (107 children, 71 adults) we detected the presence of Bet v1, Bet v2 and Bet v4 specific IgE antibodies. RESULTS: Bet v1 specific IgE antibodies were identified in most patients without any significant differences between children and adults. Bet v2 positivity was found more frequently in the group of children than in adults (p = 0.02). In most adult patients Bet v1 monospecificity was more expressed as compared to the pediatric group. More allergic subjects reacted against minor birch allergens in the pediatric group (p = 0.02). Specific IgE antibodies against Bet v1 were not detected in 10% of the tested patients. In this group, 5% of birch pollen-allergic patients were found to not have specific IgE antibodies against any of the tested recombinant allergens. CONCLUSION: The investigation of specific IgE antibodies against Bet v1, Bet v2 and Bet v4 demonstrated that the specificity of allergen-induced IgE antibodies in birch pollen-allergic individuals is dependent not only on the region in which a patient lives but also on age. Especially in children, there is an increase in the number of allergic subjects who do not react exclusively against the major allergen. The question is whether some allergen-specific IgE antibodies will disappear depending on age or on the contrary whether their synthesis will be increased.

Immunological studies on human larval toxocarosis.

The aim of the study was to characterize the antiparasite humoral response in patients with the syndrome of visceral larval toxocarosis. Specific IgG, specific IgE and total IgE immunoglobulins against Toxocara canis excretory/secretory antigens (TES) were detected by using ELISA technique. Antibody response was studied in complete sera as well as in immunoglobulin fractions (IgG and IgE), isolation of which was performed on Protein A Sepharose. It was observed that removal of IgG from the serum samples resulted mostly in increasing levels of anti-Toxocara IgE antibodies what agrees with the theory of the blocking effect of IgG in the immune response. The results demonstrated a little correlation between slgG and slgE in the sera of symptomatic patients, examined in ELISA reaction.

[Less common causes of chronic tubulointerstitial nephritis complicated by acquired Fanconi syndrome]. / Méne obvyklá prícina chronické tubulointersticiální nefritidy komplikované získaným Fanconiho syndromem.

The authors describe an uncommon form of the disease in a 25-year-old man who developed renal failure on the background of tubulointerstitial nephritis combined with acquired Fanconi syndrome. The authors analyze in detail the results of the laboratory examination and present differential diagnostic reflections which lead, after elimination of other possibilities, to the final diagnosis of immunity-mediated, viral antigen-conditioned tubulointerstitial nephritis.

[Detection of specific antibodies to all classes, including IgE, against pollen allergens using the ELISA test]. / Prukaz specifických protilátek vsech tríd, vcetne IgE, proti pylovým alergenum testem ELISA.

The authors describe a modification of the immunoenzymatic test suitable for estimation (sensitive dedection) of specific antibodies against pollen allergens of all classes of immunoglobulins, incl. specific IgE. When introducing the method, sera of experimental rabbits immunized with selected allergens were used. After elaboration of suitable conditions, the method was used to assess the presence of specific antibodies in sera of patients with the diagnosis of pollinosis. It was found that the most suitable concentration of pollen allergen combined with the solid phase for detection of specific IgE is 100 micrograms/ml, the optimal concentration of glutaraldehyde used to increase the amount of allergen combined with the solid phase by its polymerization and fixation is 0.25%. The use of the method does not call for special equipment and uses locally produced products. The method is useful for supplementing the diagnosis and its more accurate assessment under clinical conditions when biological tests cannot be used for diagnosis. The test makes monitoring during immunotherapy possible.

Progesterone and testosterone: contraceptive and immunosuppressive effects in mice.

Progestin (medroxyprogesterone acetate, trade name Depo-Provera, Upjohn, Belgium) in 6 body-weight-matched doses with those used in women, i.e., 0.05 mg/mouse, given intramuscularly at 4-5-day intervals (duration of one ovulation) induced a marked reduction of fertility with precocious atresia of Graafian follicles, minimal numbers of growing follicles and an excess of corpora lutea; the lymphoid system remained morphologically and functionally normal. A combination of progestin and androgen (testosterone isobutyrate, trade name Agovirin Depot Biotika, Czechoslovakia) in a total dose of 1 mg + 0.9 mg (body-weight-matched with that used in men) or one order higher and divided into 6 doses given at 17-day intervals, i.e. roughly one half of the duration of spermatogenesis, had no inhibitory effect on spermatogenesis, whereas higher doses damaged the lymphoid system. This damage was reflected in a markedly reduced thymus weight with great depletion of cortex lymphocytes and diminished capacity for antibody formation to sheep red blood cells. The significance of the immunosuppressive effect for estimating the risks involved in the administration of sex steroids in human contraception is discussed.

H-2 antigenicity of Leydig cells.

Partial absorption of oligospecific reagents by the particulate membrane fraction prepared from isolated interstitial cells grown in culture (the harvested cell population contained about 80% Leydig cells)suggests that membranes of Leydig cells carry antigenic specificities H-2K (11,25) and H-2D (4) but not antigens controlled by the I region. The results of absorption experiments have been confirmed by the methods at the level of individual cells; the native Leydig cells gave a positive reaction in the dye exclusion cytotoxic and immunofluorescent tests with he polyvalent regent B10D2 and B10 (directed against antigens of the regions H-2K through H-2I-E).

Spermatogenesis and immune responsiveness to sheep red blood cells in guinea pigs treated with cyproterone acetate and testosterone.

The synthetic steroid antiandrogen, cyproterone acetate, and the androgen, testosterone isobutyrate, were injected subcutaneously into adult male and female guinea pigs of two outbred strains (laboratory coloured and albino Pirbright-Harley stocks). Cyproterone acetate was given as 1, 10 and 20 daily 5-mg doses (body-weight matched with immunosuppressively effective doses in mice) and as 30 doses of 15 micrograms/100 g b.w. (body-weight matched with those used in the long-term human male contraception), testosterone as 1 and 10 doses of 5 mg/100 g b.w. The guinea pigs of the two stocks used showed different sensitivity: with the same doses of CA albino guinea pigs compared to the coloured ones declined in weight, had lower viability, reduced thymus weight, diminished capacity for SRBC haemolysin formation, a lower testes weight and inhibited spermatogenesis. Both stocks reacted by similar lymphopoenia, however. Testosterone neither inhibited haemolysin formation nor produced lymphopoenia but induced a more marked reduction of thymus weight than did CA and, in addition, produced greater depletion of cortex lymphocytes in albino guinea pigs. The thymus of albino guinea pigs was more sensitive to testosterone than to cyproterone acetate. Sexual dimorphism was apparent in some criteria in albino guinea pigs following cyproterone acetate and testosterone. Cyproterone acetate in the lowest doses had no inhibitory effect on thymus or SRBC formation but also did not affect spermatogenesis in albino guinea pigs. The hitherto accepted classification of animal species according to which guinea pigs (and also man) belong to steroid-resistant species is discussed and questioned, and the potential risks of application of sexual steroids in male contraception are stressed.

Effect of some steroid hormones on the homing affinity of lymph node cells and monocytes.

The selective homing of labelled cells from lymph nodes and peritoneal exudate to lymph nodes and spleens is strongly inhibited after an in vivo administration of steroid hormones, cyproterone acetate, hydrocortisone acetate and hydrocortisone succinate. At the same time the number of lymphocytes and monocytes is greatly reduced in the peripheral blood. This decrease and changes in recirculation of lymphocytes and monocytes are due not to the lysis of cells but to an altered distribution of them in the body and an increased trapping in the bone marrow. The values in the peripheral blood picture are very rapidly reconstituted after the level of the soluble, short-term acting steroid--hydrocortisone succinate--in the body had decreased, and the transferred labelled cells re-enter the lymph nodes while their number in bone marrow is reduced. The altered distribution of cells in the body that is obviously not advantageous for the immune reactions may be one of the causes of the immunosuppressive effect of the steroids.

Are histocompatibility antigens in neonatal skin grafts truly tolerogenic?

Neonatal skin grafts across a weak histocompatibility barrier (MSA-incompatible) were used only for sensitization of the recipients whose responsiveness was then tested in two different assays. On the basis of 1. functional inactivation of PEC in a PEC transfer system and 2. inhibition of macrophage migration. The difference between neonatal and adult skin grafts turned out to be quantitative rather than qualitative, i.e., both induced sensitization whose demonstration with the neonatal grafts required more sensitive techniques and/or more favourable timing because it was weaker. The possible nature of the difference, which was occasionally interpreted as being due to a tolerogenic rather than immunogenic activity of neonatal grafts, is discussed.

Reversible inhibitory effect of the non-steroidal antiandrogen flutamide (SCH 13521) on spermatogenesis in mice.

The effects of a prolonged subcutaneous administration of SCH 13521 dissolved in 0.3% hydroxypropyl cellulose (2-8 weeks in daily doses of 0.2 or 1.0 mg amounting to an estimated equivalent of experimental and curative doses used by others in laboratory animals and men) were studied in males of the mouse inbred strain C57BL/6. Following the treatment, the activity of spermatogenesis (expressed as the mean number of seminiferous tubules containing mature sperm and epididymal sperm count) was inhibited while the testis weight was not reduced, obviously due to an absolute increase of the interstitial tissue which was a marked histological feature of the testes, particularly following the higher doses of SCH 13521. Lower doses and shorter-lasting administration of the compound seem to inhibit the activity more effectively because after a prolonged administration reparatory processes tend to be triggered via a stimulatory effect on the synthesis of testosterone in Leydig cells. The solvent alone, hydroxypropyl cellulose, had some inhibitory effect on spermatogenesis. The lymphoid system remained both morphologically and functionally unaffected by SCH 13521 unlike the steroidal antiandrogen cyproterone actetate.

Effect of an antiandrogen on the lymphoid system.

A side-effect of the administration of cyproterone acetate, an antiandrogenic steroid, to newborn, juvenile or adult male mice (in doses comparable to those used clinically) was found in a marked reduction of the white pulp of the spleen and reduced weight or even absence of the thymus.

Analysis of different survival of weakly incompatible allografts of skin from the ear and back.

A comparison of the survival of skin grafts across the H-barrier presented by the male-specific antigen in mice was made with grafts of skin from different sources, namely from the back or from the ear. Their survival was roughly similar provided the ear skin was depleted of the subepidermal connective tissue with the cartilage. With this layer intact, the ear skin grafts had a prolonged survival and some of them even took permanently. A local administration of a mucolytic agent to the normal grafts from the back accelerated their rejection. The factors of the subepidermal connective tissue which might be responsible for the effect on the survival of weakly incompatible skin grafts and in this connection the role of the source of the skin are discussed.

Changes in agglutination of cells with H-2 antibodies or lectins and in stimulation of lymphocytes as induced by certain polymers (chondroitin sulphate/PVP/PHA/Con A/capping of H-2 antigens).

The effect of certain polymers on the course of immune interaction has been well known although its mechanism at the cellular level remains obscure. We thus affected cells in vitro with two of such substances, CS and PVP, and followed their behaviour in various assays. It was shown that in this way the agglutinability of mouse erythrocytes with H-2 antibodies or of mouse lymphocytes with threshold doses of Con A could be enhanced, antibody-induced capping of H-2 antigens on lymphocytes accelerated and PHA-induced stimulation of mouse lymphocytes inhibited. The possible mechanisms of these changes putatively mediated through the effect on the cell membrane or cell coat are discussed.

The significance of the cell surface for the homing affinity of injected lymphoid cells in mice.

51Cr-labelled lymph node cells were injected into syngeneic or non-syngeneic mice and their relative tissue distribution was compared under conditions where, for the reason of either timing or genetic combination, the host-versus-graft response could be neglected. The immigration of injected lymphoid cells to lymph nodes was markedly reduced when the recipients were non-syngeneic or when the level of glycosaminoglycans in their blood was artificially increased. In contrast, the homing affinity to the spleen was inhibited only for a relatively short period of time following stress. In the various tested situations in which the migration of cells to lymph nodes was inhibited, it seemed to be the relationship of the cell surfaces of the sessile and circulating cells which played an important role in the outcome of their interactions. This relationship may be influenced both by certain genetic differences and experimentally, for example, through adsorption.

Immune capacity of mice tolerating a weakly incompatible skin allograft. I. Tolerance induction by neonatal skin graft in adult recipients or by neonatal inoculation of adult cells.

An analysis was made of the so-called "tolergenic" effect of neonatal skin grafts which tend to survive longer than control adult grafts and confer this tendency also on simultaneously transplanted adult grafts of the same genotype. When the time interval between the neonatal and adult grafts varied from zero to 150 days, it could be demonstrated that neonatal skin grafts are capable of inducing second-set response comparable to that induced by adult grafts provided that the adult graft is not given simultaneously but only following 20 days or later. Signs of immune erises (i.e. various degrees of a round-cell infiltration) were, however, detected histologically even with shorter intervals of time. The involvement of some non-specific components in the early postgrafting period was indicated by a similar effect of sera from recipients of syngeneic grafts and MSA-incompatible grafts on the survival of grafts across the male-specific antigen barrier. Only long surviving allografts tend to specifically reduce the recipient's immune responsiveness. The possible mechanisms of the specific and non-specific components in the effect of neonatal skin grafts and the factor of time are discussed.