Modern fluorescent techniques to investigate the mechanisms of lymphocyte activation.

Fluorescent proteins are promising toolsfor studying intracellular signaling processes in lymphocytes. This brief review summarizes fluorescence-based imaging techniques developed in recent years and discusses new methodological advances, such as fluorescent photoswitches, fluorescence recovery after photobleaching (FRAP), fluorescent resonance energy transfer (FRET), fluorescence lifetime imaging microscopy (FLIM), photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), stimulated emission depletion (STED), total internal reflection fluorescence (TIRF) and other techiques. This survey also highlights recent advances in vitro imaging of live tissues, novel applications of flow cytometry with genetically modifed fluorescent proteins, and future prospects for the development of new immunological test systems based on fluorescent protein technology.

[Transfer factor of immune reactivity to diphtheria-tetanus anatoxin modulates the action of neurotransmitters in the intestinal smooth muscle].

Transfer factor (TF) of immune reactivity (10(-5) - 10(-3) mg/ml) to diphtheria-tetanus anatoxin modulates slow waves and spontaneous contractile activity of non-atropinized smooth muscle stripes (SMS) of guinea-pig taenia coli. TF (10(-4) mg/ml) transforms slow waves into stable depolarization and tonic contraction. After SMS atropinization, the substance acts in the same way. In the presence of methylene blue (10(-5) M), a guanylatecyclase blocker, FT induces transitory increase of SMS muscle tone, which is followed by their stable relaxation. ATP and UTP, purinoceptors agonists, evoke substantial hyperpolarization of smooth muscle cells membrane and their relaxation. FT enhances post-inhibitory excitation in SMS. In the presence of acetylcholine (10(-5) M) FT (10(-4) mg/ml) transforms the inhibitory ATP action on tonic contraction into excitative. This substance (10(-5), 10(-4) mg/ml) enhances Ca2+ mobilization from ryanodine-sensitive calcium store, inhibits the release of these cations from IP3-sensitive calcium store of sarcoplasmic reticulum. TF demolishes the inhibitory actions of sodium nitroprusside (nitric oxide donor), and noradrenaline in taenia coli smooth muscles.

[Effects of Staphylococcus aureus cell-bound protein A on adenosine triphosphate and nitric oxide inhibitory actions in smooth muscles].

Cell-bound protein A (CBPA), an immune-active substance of Staphylococcus aureus was ascertained to depolarize membrane of taenia coli smooth muscle (SM) cells, depress ATP inhibiting action (or uridinetriphosphate (UTP)) and sodium nitroprusside (SNP). ATP or UTP-induced membrane hyperpolarization increased during first minutes of CBPA exposure. Bacterial substance enhanced and then inhibited fast component of nicotine-induced relaxation of histamine-activated smooth muscles. This enhancement was inhibited by N(omega)-nitro-Larginine, a NO-syntase blocker. CBPA decreased ATP inhibiting action upon histamine-induced contraction, but enhanced cholinergic SM excitation. All these processes are reversible.

[Transfer factor modulates inhibitory action of neurotransmitters on intestinal smooth muscles]. / Faktor perenosu moduliuie hal'mivnu diiu neiromediatoriv na hladen'ki m'iazy kyshechnyka.

It has been shown that Transfer factor (TF) to Staphilococcus aureus antigens blocked ATP-induced component of the inhibitory junction potential in taenia coli smooth muscle from guinea-pig, and converted an inhibitory action of exogenous ATP into an exciting one (instead of the hyperpolarization of the smooth muscle, TF induced its depolarization). TF at 10-6 mg/ml converted the relaxing effects of sodium nitroprusside (nitric oxide donor) into exciting ones in smooth muscle strips. Higher concentrations (10-5-10-3 mg/ml) of TF slightly amplified the relaxing effect of sodium nitroprusside. Both a- and b-adrenergic activation in taenia coli smooth muscle were not sensitive to that agent.

[Effect of staphylococcus active substances on ATPase activity of smooth muscle actomyosin and myosin]. / Vpliv aktyvnykh substantsii stafilokoka na ATP-aznu aktyvnist' aktomiozynu ta miozynu hladen'kykh m'iaziv.

The effect of staphylococcus active substances--protein A (PA) and peptidoglican (PG) at concentrations 10(-6)-10(-2) mg/ml on the ATPase activity of pig stomach natural actomyosin and myosin was studied. It was shown that PA and PG at direct contact with smooth muscle contractile proteins caused the activation and inhibition of ATPase activity, respectively. On the basis of this investigation it was assumed that staphylococcal active substances were able to modify of the ATPase activity smooth muscle contractile proteins perhaps via direct action on the myosin molecule, which could be accompanied by conformational changes of the active center of myosin ATPase.

[The immune reactivity transfer factor as a modulator of lymphocyte functional activity in rats]. / Faktor perenosu imunnoï reaktyvnosti iak moduliator funktsional'noï aktyvnosti limfotsytiv shchuriv.

The transplantation of the thyroid tissue is one of the perspective methods for rehabilitation of thyroid gland functional disorders that appear due to the influence of insufficient environmental conditions on organism. By means of micromethod of lymphocyte blast transformation reaction on the base of [3H]-thymidine shift the functional activity of the Wistar rat's splenocytes was studied in case of radiation induced hypothyroidism with or without xenotransplantation of newborn pig thyroid gland organ culture. It was found that the level of thyroxine and triiodothyronine significantly decreased in serum of irradiated animals, the lymphocyte proliferation level was also reduced (by means of radioiodine introduction in dose of 8,325 MBk/mmole). Application of thyroid gland tissue xenotransplantate in this model of hypothyroidism helped to achieve the increasing of thyroid hormones levels in serum and rehabilitation of lymphocytes functional activity. The opportunities for correction of immunological disorders with the help of transfer factor of immune reactivity preparates were investigated. Transfer factor--is a low-molecular weight leukocyte extract (J 10 kD) with immunomodulating activities. This preparates activated the proliferation of splenocytes from animals with hypothyroidism and animals with hypothyroidism after xenotransplantation.

[The role of microorganisms of the wound in postop complications]. / Rol' mikroorhanizmiv rany u vynyknenni pisliaoperatsiinykh uskladnen'.

Sensitivity of mostly wide-spread pathogenic microorganisms to modern antibiotics of the wide spectrum of action (Tienam, Mefoxin, Cifran), and to antibiotics, widely applied in everyday clinical practice was studied.

[Mitogenic effects of staphylococcal antigenic substances on irradiated lymphocytes from Wistar rats]. / Mitohenna dia antyhennykh substantsii stafilokoka na limfotsyty oprominenykh shchuriv liniï vistar.

Proliferation activity of lymphocytes from rats was studied using a micromethod of blasttransformation reaction after X-ray irradiation (absorbed doses for animals were 0.5 and 1 Gy). Staphylococcus protein A and peptidoglycan stimulated the proliferation of irradiated lymphocytes. It was assumed that staphylococcus antigen substances held radiation protective effects. Staphylococcus protein A may be used for stabilization of lymphopoesis and functional activity of humoral and cellular immunity in irradiated animals due to T- and B-mitogenic properties.

[DNA synthesis in lymphocytes of irradiated animals as affected by mitogens and Staphylococcal antigens]. / Syntez DNK u limfotsytakh oprominenykh tvaryn pid vplyvom mitoheniv i antyheniv stafilokoka.

Proliferation activity of lymphocytes from rats was studied using micromethod of blast transformation reaction after exposure of X-ray irradiation (absorbed doses for animals are 0.5 and 1 Gr). The reducing of the functional activity of lymphocytes was found. There was difference in lymphocyte response induced by standard mitogens (PHA, ConA, LPS) and antigens of Staphylococcus protein A and peptidoglycan. The irradiation protective effects of Staphylococcus substances were found.

[Human specific transfer factor to Staphylococcus aureus antigens]. / Liuds'kyi spetsyfichnyi Faktor perenosu do antyheniv Staphylococcus aureus.

Immunobiological properties of human specific transfer factor (TF) to Staphylococcus aureus antigens were studied. It is shown that this TF activated human leucocytes in vitro as well as in vivo. Antigen specificity of TF's immunomodulating effects is also shown. In vitro we used leucocyte migration inhibition test (IML), macrophage inhibition test (MPI) and rosette formation (E-ros). For testing in vivo we used delayed type hypersensitivity (DTH) skin tests.

[The biological activity of the transfer factor induced by bacterial antigens]. / Biolohichna aktyvnist' faktora perenosu, indukovanoho bakterial'nymy antyhenamy.

Today's statement of transfer factor, an immunostimulator derived from leukocytes which enhances antiinfectious immunity, is observed in the review. Basic biological, physical and chemical characteristics of the transfer factor, its possible action mechanisms, and laboratory and clinical methods of use to cure infectious fungal (Candida, Coccidium), invasive (schistosomiasis, leishmaniasis, cryptosporidiosis), viral (varicella zoster, ophthalmic herpes, Herpes simplex types 1 and 2, H. zoster, H. simplex ceratitis, genital herpes, human herpes virus type 6, postherpetic neuritis, hepatitis B, AIDS), and bacterial infections (Mycobacterium leprae, M. tuberculosis, M. fortuitum, Salmonella cholerae suis, S. dublin, S. Virchov, Brucella abortus, Actinobacillus pleuropneumoniae, bacterial sepsis, Staphylococcus) are described.

[The isolation of a transfer factor of the delayed-hypersensitivity type to the antigenic substances of Staphylococcus aureus in guinea pigs]. / Oderzhannia faktora perenosu hiperchutlyvosti spovil'nenoho typu mors'kykh svynok do antyhennykh substantsiï Staphylococcus aureus.

Immunoactivating effects of guinea pig's staphylococcal Transfer factor (TF) were studied in homologous and heterologous (human leukocytes) systems. In vivo (skin tests) and in vitro (Inhibition of leukocytes migration) tests showed that our TF is able to activate intact as well as sensibilized cells. Antigen-specific properties of TF also were showed.