The bovine dialysable leukocyte extract IMMUNEPOTENT CRP induces immunogenic cell death in breast cancer cells leading to long-term antitumour memory.

BACKGROUND: Cancer recurrence is a serious problem in breast cancer (BC) patients, and immunogenic cell death (ICD) has been proposed as a strategy to overcome this recurrence. IMMUNEPOTENT CRP (ICRP) acts as an immunomodulator and can be cytotoxic to cancer cells. Thus, we evaluated if ICRP induces ICD in BC cells. METHODS: Immunogenicity of ICRP-induced cell death was evaluated in vitro, analysing the principal biochemical characteristics of ICD in MCF-7, MDA-MB-231 and 4T1 cells. Ex vivo, we assessed the ability of killed cancer cells (KCC) obtained from ICRP-treated 4T1 cells (ICRP-KCC) to induce DC maturation, T-cell priming and T-cell-mediated cancer cytotoxicity. In vivo, we evaluated tumour establishment and antitumour immune memory after prophylactic ICRP-KCC vaccination in BALB/c mice. RESULTS: ICRP induced caspase-independent, ROS-dependent cell death, autophagosome formation, P-eIF2α, chaperone protein exposure, CD47 loss, ATP and HMBG1 release in BC cells. Additionally, ICRP-KCC promoted DC maturation, which triggered T-cell priming and cancer cytotoxicity. Prophylactic vaccination with ICRP-KCC prevented tumour establishment and induced long-term antitumour memory in BALB/c mice, involving DC maturation in lymph nodes, CD8+ T-cell augmentation in lymph nodes, peripheral blood and tumour site and ex vivo tumour-specific cytotoxicity by splenocytes. CONCLUSIONS: ICRP induces ICD in BC cells, leading to long-term antitumour memory.

IMMUNEPOTENT CRP induces DAMPS release and ROS-dependent autophagosome formation in HeLa and MCF-7 cells.

BACKGROUND: IMMUNEPOTENT CRP (ICRP) can be cytotoxic to cancer cell lines. However, its widespread use in cancer patients has been limited by the absence of conclusive data on the molecular mechanism of its action. Here, we evaluated the mechanism of cell death induced by ICRP in HeLa and MCF-7 cells. METHODS: Cell death, cell cycle, mitochondrial membrane potential and ROS production were evaluated in HeLa and MCF-7 cell lines after ICRP treatment. Caspase-dependence and ROS-dependence were evaluated using QVD.oph and NAC pre-treatment in cell death analysis. DAMPs release, ER stress (eIF2-α phosphorylation) and autophagosome formation were analyzed as well. Additionally, the role of autophagosomes in cell death induced by ICRP was evaluated using SP-1 pre-treatment in cell death in HeLa and MCF-7 cells. RESULTS: ICRP induces cell death, reaching CC50 at 1.25 U/mL and 1.5 U/mL in HeLa and MCF-7 cells, respectively. Loss of mitochondrial membrane potential, ROS production and cell cycle arrest were observed after ICRP CC50 treatment in both cell lines, inducing the same mechanism, a type of cell death independent of caspases, relying on ROS production. Additionally, ICRP-induced cell death involves features of immunogenic cell death such as P-eIF2α and CRT exposure, as well as, ATP and HMGB1 release. Furthermore, ICRP induces ROS-dependent autophagosome formation that acts as a pro-survival mechanism. CONCLUSIONS: ICRP induces a non-apoptotic cell death that requires an oxidative stress to take place, involving mitochondrial damage, ROS-dependent autophagosome formation, ER stress and DAMPs' release. These data indicate that ICRP could work together with classic apoptotic inductors to attack cancer cells from different mechanisms, and that ICRP-induced cell death might activate an immune response against cancer cells.

Quality consistency evaluation of commercial transfer factor injections by chromatographic fingerprint combined with multivariate statistical analysis.

The current quality control methods relying mainly on chromogenic reaction can hardly ensure the quality and safety of the biochemical drug with complex chemical composition. Therefore, a chromatographic fingerprint method was developed for the quality evaluation of a multicomponent biochemical drug, transfer factor injection. High-performance liquid chromatography fingerprint was measured by using a C18 column (250 × 4.6 mm, 5 µm) with a mobile phase composed of 0.1% trifluoroacetic acid-water and 0.085% trifluoroacetic acid-acetonitrile under gradient elution. The developed method was validated and was subsequently applied to 57 batches of commercial products which were sampled by National Drug Assessment Program. High-resolution mass spectrometry analysis was performed on characteristic peaks of fingerprints, and a series of amino acids, nucleosides, and deoxynucleosides were identified. In the fingerprint assessments, principal component analysis and Hotelling T2 analysis yielded the best results. The results generally indicated that there was a significant difference among products of batch-to-batch or from different manufacturers. Abnormal samples and its discriminatory components were also explored. In summary, the established fingerprinting method with multivariate statistical analysis could offer an efficient, reliable, and practical approach for quality consistency evaluation of transfer factor injection, providing a reference for the quality control of other multicomponent biochemical drugs.

The utilisation of human dialyzable leukocyte extract (IMMODIN) as adjuvant in albendazole therapy on mouse model of larval cestode infection: Immunomodulatory and hepatoprotective effects.

Metacestode (larval) stages of zoonotic cestodes of medical and veterinary importance cause chronic infections associated with immunosuppression. During mouse model of cestode infection induced by larvae of Mesocestoides (M.) vogae, we investigated the effects of dialyzable leukocyte extract (DLE) containing low-molecular weight substances (under 10 kDa) prepared from peripheral blood leukocytes of healthy human donors (available under commercial name IMMODIN). In the experiment, the effects of DLE as adjuvant to anthelmintic albendazole (ABZ) as well ABZ mono-therapy were also investigated. We showed that DLE enhanced therapeutic effect of ABZ by significant reduction of parasites number in both biased sites. Furthermore, administration of DLE reduced fibrosis and concentrations of lipid peroxides in the liver and thereby showed cytoprotective effect. In contrast, higher hydroxyproline level and numbers of larvae enclosed in fibrous capsules were found in ABZ-treated group. In order to investigate whether DLE could affect parasite-induced immunosuppression, we evaluated selected immune parameters. The results showed that DLE administration to mice increased proliferation of concanavalin A stimulated splenic cells ex vivo. Similarly, in vitro study confirmed that DLE ameliorated hypo-responsiveness of T lymphocytes and partially reverted suppressive effect of parasites excretory-secretory products. In addition, flow cytometric analysis revealed higher numbers of T helper and NK cells in the spleen and peritoneal cavity of infected mice after DLE + ABZ therapy. We also found strongly reduced serum levels of TGF-ß1 and IL-17 as well as modulation of cytokines associated with Th1/Th2 immunity. These results suggest that IMMODIN could serve as a suitable adjuvant to the primary anthelmintic therapy.

[Reactivation of patch-stage mycosis fungoides caused by oral administration of dialyzable leukocyte extract. Case report]. / Reactivación de micosis fungoide en estadio de placa por administración de extracto dializado de leucocitos vía oral. Reporte de caso.

BACKGROUND: Mycosis fungoides is a cutaneous T-cell lymphoma. The patch stage is limited to the skin and may spontaneously involute or progress, spreading to peripheral blood, lymph nodes and viscera. CASE REPORT: 64 year-old female with a 6-year history of dermatosis with scaly, poorly delimited and pruritic plaques on the chest and extremities. She had received oral steroids and antihistamines, with transient partial remissions been experienced. Skin biopsy revealed Pautrier's microabscesses, which are pathognomonic of mycosis fungoides. Positron-emission tomography and peripheral blood smear ruled out dissemination and confirmed patch-stage mycosis fungoides. She received nitrogen mustard topical derivatives, psoralen plus UVA therapy, steroids and tacrolimus. She achieved complete remission at 6 months. Two years later, she was treated with dialyzable leukocyte extract, which reactivated the patch lesions with severe itching; the extract was discontinued. The lesions resolved two weeks after topical clobetasol was applied. CONCLUSIONS: Th2 predominates in mycosis fungoides. Given that dialyzable leukocyte extract reinforces the Th1 profile, it was unlikely for it to reactivate the disease, but the diversity of lymphocyte immunophenotypes in mycosis fungoides and the complex activation networks caused a paradoxical reactivation.

Antecedentes: La micosis fungoide es un linfoma cutáneo de células T. El estadio de placa se encuentra limitado a piel y puede involucionar o progresar, diseminándose a sangre periférica, ganglios y vísceras. Reporte de caso: Mujer de 64 años de edad con dermatosis de seis años de evolución con placas descamativas, mal delimitadas y pruriginosas en tórax y extremidades. Había recibido esteroides orales y antihistamínicos, con los que presentaba remisiones parciales transitorias. Mediante biopsia cutánea se encontraron microabscesos de Pautrier, patognomónicos de micosis fungoide. La tomografía por emisión de positrones y el frotis de sangre periférica descartaron diseminación y confirmaron micosis fungoide en estadio de placa. La paciente recibió derivados tópicos de mostaza nitrogenada, psoralenos con radiaciones ultravioleta A, esteroides y tacrolimus. Presentó remisión total a los seis meses. Dos años después recibió extracto dializado de leucocitos, con el que se reactivaron las lesiones con prurito intenso; suspendió el extracto. Las lesiones involucionaron dos semanas después de iniciar el clobetasol tópico. Conclusiones: En la micosis fungoide predomina Th2. Dado que el extracto dializado de leucocitos refuerza el perfil Th1 no se esperaba que reactivara la enfermedad, pero los diversos inmunofenotipos de los linfocitos en la micosis fungoide y las complejas redes de activación ocasionaron reactivación paradójica.

[Effect of Acupoint Injection on Eosinophil Counts,Protein and mRNA Expressions of Eotaxin in Nasal Mucosa of Allergic Rhinitis Rats].

OBJECTIVE: To observe the effect of acupoint injection on eosinophils (EOS) counts and expression of eotaxin in nasal mucosa of allergic rhinitis (AR) rats, so as to reveal its mechanism underlying improving AR. METHODS: Twenty-four Sprague-Dawley (SD) rats were randomly divided into normal, model and acupoint injection groups (n=8 in each group). The AR model was established by intraperitoneal injection of ovalbumin sensitization. Bilateral "Yingxiang"(LI 20) and "Yintang"(GV 29) were selected for acupoint injection of the mixture solution of lidocaine, dexamethasone, and transfer factor (0.1 mL/acupoint) on the 1st, 5th, 9th, and 13th day after AR model established, a total of four times. EOS in the nasal mucosa was counted under light microscope after HE staining. Protein and mRNA expressions of eotaxin in the nasal mucosa were detected by immunohistochemical and RT-PCR methods, respectively. RESULTS: Compared with the normal group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly higher in the model group (P<0.05). Compared with the model group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly lower in the acupoint injection group (P<0.05). CONCLUSIONS: Acupoint injection can reduce the nasal mucosa inflammation by suppressing the protein and mRNA expressions of eotaxin, decreasing the infiltration and gathering of EOS in the nasal mucosa.

Serum Derived Transfer Factor Stimulates the Innate Immune System to Improve Survival Traits in High Risk Pathogen Scenarios.

Preclinical Research Transfer Factors (TFs) are low molecular weight (<5,000 daltons) biological response mediators. In the present study, a serum derived TF improved the ability of the recipient animal to survive high-risk infectious challenges (salmonellosis and canine parvoviral enteritis (CPV)) by altering the host's cytokine response profile. Mice mortally challenged with 5,000 colony-forming units of Salmonella experienced a group mortality of 73% while mice treated with a single 5 mg dose of the TF demonstrated a significant decrease in morbidity (7%, p ≤ 0.01). The splenic bacterial load in untreated mice was over 10,000 times higher than that in the TF treated mice. Twenty-four hours post-administration, the treated murine population expressed a rapid temporal increase in serum IL-6 (26-fold) and INF-γ (77-fold) concentrations. IL-6 can act as a critical signal regulating action against bacterial pathogens. A comparative double-blind study performed using dogs confirmed to be undergoing a canine parvovirus challenge showed that when conventional supportive therapy was supplemented with a single 5 mg TF dose there was a reduction (p ≤ 0.01) in group mortality (68% of the TF treated group survived versus 32% of the placebo group), an observation consistent with the observed increase in INF-γ, a cytokine associated with promoting antiviral activity. Drug Dev Res 78 : 189-195, 2017. © 2017 Wiley Periodicals, Inc.

Anti-Inflammatory Effect of Dialyzable Leukocyte Extract in Autoimmune Prostatitis: Evaluation in Animal Model.

Objective. To evaluate the anti-inflammatory properties of Dialyzable Leukocyte Extract (DLE) in a murine model of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Methods. Histopathological characterization, prostatein Enzyme-Linked Immunosorbent Assay, and immunohistochemical analysis for CD45, TNF-α, IFN-γ, IL-6, IL-17, and IL-4 molecules were done in prostatic Wistar rats treated with DLE, placebo, or Dexamethasone. Results. Histopathological analysis of animals induced to prostatitis showed inflammatory infiltrate, mainly constituted by leucocytes and mast cells as well as Benign Prostatic Hyperplasia. Serum prostatein concentrations were 14 times higher than those displayed by healthy animals. After DLE and Dexamethasone treatments, the inflammatory infiltrate decreased; the tissue morphology was similar to that of a normal prostate, and the prostatein decreased to the basal levels of healthy animals. DLE treatment produced a decreased expression of the cell surface marker CD45 and the proinflammatory cytokines TNF-α, IFN-γ, IL-6, and IL-17. On the other hand, the expression of anti-inflammatory cytokine IL-4 increased in both the Dexamethasone and DLE groups. Conclusion. DLE is able to modulate the inflammatory response in Experimental Autoimmune Prostatitis (EAP).

[Analysis on Influencing Factors of Acupoint Injection of Drugs for Improving Allergic Rhinitis in Rats].

OBJECTIVE: To observe the therapeutic effect of acupoint injection of mixture solution of lidocaine, dexa-methasone (DX) and transfer factor on symptoms and serum histamine contents in allergic rhinitis (AR) rats, so as to differentiate the role of acupoint stimulation or drug factor influencing the therapeutic effect. METHODS: SD rats were randomized into normal (n=8), model, non-acupoint-drug injection, acupoint-drug injection and acupoint-saline injection groups (n=11 in each group). The AR model was established by intraperitoneal injection of suspension of ovalbumin (0.3 mg) and aluminum hydroxide (30 mg), 1 mL, once every other day for 7 days, followed by nose-dripping of 5% ovalbumin (50 µL) for other 7 days. The AR scores for rhinocnesmus severity (1-2 points), sneezing times (1-3 points) and nasal discharge quantity (1-3 points) were assessed by cumulative quantification scoring method. "Yintang" (EX-HN 3) and bilateral "Yingxiang" (LI 20) were used for acupoint injection of mixture solution of 1% Lidocaine, DX and transfer factor (1:1:1, 0.1 mL/acupoint), once every other day for 7 days. The non-acupoints were located at the bilateral sub-costal regions and the mid-point between the left "Houhai" (GV 1) and "Huantiao" (GB 30).The same dose of saline was injected into the aforementioned acupoints for control. Serum histamine content was detected by using ELISA, and the nasal mucosal pathological changes were also observed after H.E. stain. RESULTS: After the treatment, AR modeling induced increase of symptom scores and serum histamine contents were significantly decreased in both acupoint-saline injection and acupoint-drug injection groups (P<0.01) but not in the non-acupoint-drug injection group (P>0.05), and the effects of acupoint-drug injection were markedly superior to those of non-acupoint-drug injection group (P<0.01). CONCLUSIONS: Acupoint injection of mixture solution of Lidocaine, DX and transfer factor is effective in relieving the allergic symptoms and reducing the nasal mucosa inflammation in AR rats, which may be related to its action in suppressing histamine releasing, mainly by acupoint stimulation. It suggests that the therapeutic effect mainly derives from acupoint-injection stimulation, then the injected drugs.

[Preparation and immunological evaluation of oral solution of egg yolk-derived hepatitis B virus-specific transfer factor].

OBJECTIVE: To prepare the oral solution of egg yolk hepatitis B virus (HBV)-specific transfer factor (EYHBV-TF) and evaluate its immunological activity as an immune regulator against hepatitis B. METHODS: From hens immunized with the Hepatitis B vaccine the egg yolk was isolated to extract the specific transfer factor EYHBV-TF, and its physicochemical properties were examined. Leukocyte adhesion inhibition test (LAI) was performed to detect the immunogenic activity of EYHBV-TF. The solution of EYHBV-TF was then administered orally in normal mice, and the specific cellular immune activity induced was assayed with delayed type skin hypersensitivity test (DTH), with the non-specific immune activity assessed with immune organ index. The immune responses induced by oral EYHBV-STF solution were compared with those by EYHBV-STF injection and by different dosages (injection and oral) of porcine spleen HBV-specific transfer factor (PSHBV-STF), porcine spleen nonspecific transfer factor, and egg yolk extracts from non-immunized hens. RESULTS: The prepared EYHBV-STF oral solution, which met the standards for biological products, could inhibit leukocyte adhesion in vitro and significantly enhance mouse foot pad swelling, demonstrating its capability of transferring antigen-specific delayed type hypersensitivity reactions to naive recipient. EYHBV-STF oral solution also significantly improved the immune organ index in mice (P<0 01) with similar effects to those caused by EYHBV-STF injections and by PSHBV-STF injection and oral solution. CONCLUSION: Orally administered EYHBV-STF and EYHBV-STF injection both possess hepatitis B antigen-specific cellular immune activity and can significantly enhance specific cellular immune responses.

Enhancing immune responses to inactivated porcine parvovirus oil emulsion vaccine by co-inoculating porcine transfer factor in mice.

Inactivated porcine parvovirus (PPV) vaccines are available commercially and widely used in the breeding herds. However, inactivated PPV vaccines have deficiencies in induction of specific cellular immune response. Transfer factor (TF) is a material that obtained from the leukocytes, and is a novel immune-stimulatory reagent that as a modulator of the immune system. In this study, the immunogenicity of PPV oil emulsion vaccine and the immuno-regulatory activities of TF were investigated. The inactivated PPV oil emulsion vaccines with or without TF were inoculated into BALB/c mice by subcutaneous injection. Then humoral and cellular immune responses were evaluated by indirect enzyme-linked immunosorbent assays (ELISA), fluorescence-activated cell sorter analyses (FACS). The results showed that the PPV specific immune responses could be evoked in mice by inoculating with PPV oil emulsion vaccine alone or by co-inoculation with TF. The cellular immune response levels in the co-inoculation groups were higher than those groups receiving the PPV oil emulsion vaccine alone, with the phenomena of higher level of IFN-γ, a little IL-6 and a trace of IL-4 in serum, and a vigorous T-cell response. However, there was no significant difference in antibody titers between TF synergy inactivated vaccine and the inactivated vaccine group (P>0.05). In conclusion, these results suggest that TF possess better cellular immune-enhancing capability and would be exploited into an effective immune-adjuvant for inactivated vaccines.

[Potential of prevention of metastasizing with the aid of tumor-specific transfer-factor].

The report discusses our experimental data in support of biotherapy which uses chemotherapy and antitumor immune treatment with in vivo xenogenic transfer-factor polypeptides (TFP) isolated from lymphocytes sensitized to antigens of given tumor. After excision of primary tumor--lung carcinoma of Lewis--mice C57BL/6 were injected intraperitoneally with xenogenic TFP (200 pg/body, twice) and a cytostaic dose of cyclophosphamide. Such adjuvant chemotherapy was found to prevent metastases from spreading to the lung in 100%. The marked anti-metastatic effect of the treatment correlated with recovery of splenic cell mass and its cellular structure, higher levels of large granular lymphocytes in peripheral blood and enhanced functional activity of cytotoxic cells in vitro. Our results point to a possibility of raising efficacy of treating solid malignancies with adjuvant chemotherapy in combination with adoptive immune therapy.

[Effect of transfer factor on treatment with glucocorticoids in a group of pediatric patients with persistent moderate allergic asthma]. / Efecto del factor de transferencia en el tratamiento con glucocorticoides en un grupo de pacientes pediátricos con asma alérgica moderada persistente.

BACKGROUND: Inhaled glucocorticoids are the most effective and potent drugs used to control the inflammatory bronchial reaction in patients with asthma. There are several research projects evaluating the use of immune modulators in the treatment of the asthma related inflammatory process. OBJECTIVE: To evaluate the effect of transfer factor in the treatment of pediatric patients with moderate persistent allergic asthma in terms of inhaled glucocorticoid dosing and time of using. PATIENTS AND METHODS: Randomized, double blind, placebo controlled pilot clinical trial in a cohort of pediatric patients (6-17 years old) with moderate persistent allergic asthma. Two groups were formed. Group one received transfer factor and group two was given placebo. Both groups received conventional therapy with inhaled budesonide and formoterol. Daily respiratory symptoms (cough during day, or at night, and wheezing episodes) were recorded in a personal diary. Spirometric evaluations were performed before enrolling patients, and at 1, 3 and 6 months after. RESULTS: Eleven patients were enrolled in each group. Patients in the transfer factor group showed a statistical significant reduction in the inhaled glucocorticoid doping since month 3, and this difference was maintained until the end of study. Patients on TF group showed also a non statistical significant improvement in spirometrical findings and also showed a better asthma control. CONCLUSIONS: Transfer factor helps to reduce inhaled glucocorticoids dose in patients with allergic rhinitis; however, studies with a larger number of patients should be done in order to obtain better results.

Indications, usage, and dosage of the transfer factor.

The transfer factor (TF) was described in 1955 by S. Lawrence. In 1992 Kirkpatrick characterized the specific TF at molecular level. The TF is constituted by a group of numerous molecules, of low molecular weight, from 1.0 to 6.0 kDa. The 5 kDa fraction corresponds to the TF specific to antigens. There are a number of publications about the clinical indications of the TF for diverse diseases, in particular those where the cellular immune response is compromised or in those where there is a deficient regulation of the immune response. In this article we present our clinical and basic experiences, especially regarding the indications, usage and dosage of the TF. Our group demonstrated that the TF increases the expression of IFN-gamma and RANTES, while decreases the expression of osteopontine. Using animal models we have worked with M. tuberculosis, and with a model of glioma with good therapeutic results. In the clinical setting we have worked with herpes zoster, herpes simplex type I, herpetic keratitis, atopic dermatitis, osteosarcoma, tuberculosis, asthma, post-herpetic neuritis, anergic coccidioidomycosis, leishmaniasis, toxoplasmosis, mucocutaneous candidiasis, pediatric infections produced by diverse pathogen germs, sinusitis, pharyngitis, and otits media. All of these diseases were studied through protocols which main goals were to study the therapeutic effects of the TF, and to establish in a systematic way diverse dosage schema and time for treatment to guide the prescription of the TF.

Transfer factors as immunotherapy and supplement of chemotherapy in experimental pulmonary tuberculosis.

Problems of logistics, compliance and drug resistance point to an urgent need for immunotherapeutic strategies capable of shortening the current six month antibiotic regimens used to treat tuberculosis. One potential immunotherapeutic agent is transfer factors. Transfer factors (TF) are low molecular weight dialysable products from immune cells which transmit the ability to express delayed-type hypersensitivity (DTH) and cell mediated immunity from sensitized donors to nonimmune recipients. In this study we determined the efficiency of TF as immunotherapy to treat experimental tuberculosis. When BALB/c mice are infected via the trachea with Mycobacterium tuberculosis H37Rv there is an initial phase of partial resistance dominated by Th-1 type cytokines plus tumour necrosis factor-alpha (TNFalpha) and the inducible isoform of nitric oxide synthase (iNOS), followed by a phase of progressive disease characterized by increasing expression of IL-4, diminished expression of TNFalpha and iNOS, and low DTH. Animals in this late progressive phase of the disease (day 60) were treated with different doses of TF (one injection per week) obtained from spleen cells when the peak of immune protection in this animal model is reached (day 21), or with different doses of TF from peripheral leucocytes of PPD + healthy subjects. We show here that the treatment with murine or human TF restored the expression of Th-1 cytokines, TNFalpha and iNOS provoking inhibition of bacterial proliferation and significant increase of DTH and survival. This beneficial effect was dose dependent. Interestingly, murine TF in combination with conventional chemotherapy had a synergistic effect producing significant faster elimination of lung bacteria loads than chemotherapy alone.

[Improving treatment outcome by LeukoNorm Cytochemia in patients with multiple, failed IVF or ICSI treatment cycles]. / Verbesserung der Behandlungsergebnisse durch LeukoNorm Cytochemia bei Patientinnen mit mehrfachen, frustranen IVF- oder ICSI-Behandlungszyklen.

OBJECTIVE: The aim of these studies was to investigate whether the leucocytic ultrafiltrate LeukoNorm Cytochemia, which is approved for the treatment of immunologically-based recurrent spontaneous abortions (RSA), improves treatment results in patients with repetitive IVF or ICSI failures. MATERIAL AND METHODS: Included in this study were patients with 3 embryo transfers, with at least 8 morphologically good-looking embryos of the categories a and b, and no conception. Excluded were patients with accompanying endocrine disorders, uterine malformations, and those exhibiting the presence of an antiphospholipid syndrome and of further autoantibody syndromes (e.g. antinuclear antibodies [ANA]). The patients were treated in three prospective, randomized studies. The studies differed with respect to the frequency and timing of the administration of LeukoNorm Cytochemia. RESULTS: Pregnancy rates in each of the study protocols were generally higher than in the normal treatment group. Because of the small number of patients (study I: 10 patients), the differences were not always significant. The results of study III, which is still ongoing and in which LeukoNorm Cytochemia has been given on 5 consecutive days starting with the day of oocyte retrieval, are more significant. In the treatment group of study III the pregnancy rate was 55% as opposed to a rate of 21.2% in the non-treatment group. CONCLUSIONS: The administration of LeukoNorm Cytochemia can significantly improve treatment results in patients with repetitive IVF or ICSI failures. Currently the most favourable results are observed with a dosage of 1 unit/10 kg on 5 consecutive days, starting with the day of oocyte retrieval. These results imply that, in the IVF or ICSI programmes, there exists a group of patients with disturbances in the embryo-maternal dialogue, and therefore no conceptions. Furthermore, the results demonstrate that the administration of LeukoNorm Cytochemia can improve the implantation rate of transferred embryos in these patients. We conclude that growth factors and cytokines synthesized and secreted by leucocytes have an important influence on embryonic implantation and growth.

Clinical study on acupuncture treatment of stomach carcinoma pain.

Clinical observation on 48 cases of stomach carcinoma pain indicated that acupuncture including filiform needle group and point-injection group had better therapeutic effects in treatment of stomach carcinoma pain when patient's mind was concentrated at the site of disease. After treatment for 2 months, the long-term effective rates of analgesia in both the filiform needle group and the point-injection group were similar to that in the western medicine group, all being about 81%. While the long-term markedly effective rates in the two groups were superior to that in the western medicine group. Life quality of the patients in all the groups were improved. The toxic action and side effects caused by chemotherapy were prevented, the high viscous state showed by indexes of blood rheology was improved, and the lowered Cu-Zu-SOD activity in erythrocytes in patients of stomach carcinoma was increased in the filiform needle group and the point-injection group. Based on the results of clinical study, we consider that acupuncture analgesic effect on stomach carcinoma is related to the increase of PLEK, improvement of cellular immune function and the elevation of life quality after acupuncture.

[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.

Activities and characteristics of transfer factors.

This report summarizes three components of our transfer factor research program. Several clinical studies have used oral administration of transfer factor containing materials. Sceptics have rejected these findings by assuming that the acidic and enzymatic environment of the gastrointestinal tract would destroy the factors. To further examine this issue, we have conducted dose-response studies of the delayed-type hypersensitivity reaction in mice that were given transfer factor either by gavage or subcutaneously. There were no difference in the responses that were related to the route of administration. We conclude that oral route of administration is efficacious and should be used when possible. We have also studied the effects of transfer factors on immune responses by recipients. The details of this research are presented in the paper by Dr. Alvarez-Thull. Briefly, the study showed that recipients of a specific transfer factor responded to the antigen for which the factor was specific by secreting gamma-IFN, but no other cytokines. The structures of transfer factor molecules are unknown. We have developed a process for isolating transfer factors in pure form and we have obtained preliminary data concerning amino acid sequences. Our goal is to obtain the complete primary structure of several transfer factor molecules.

Murine transfer factors: dose-response relationships and routes of administration.

Transfer factors are protein immunomodulators that transfer the ability to express cell-mediated immunity from immunized donors to nonimmune recipients. The effects are antigen-specific. The experiments described in this report are a comparison of the relationship of the route of administration of various transfer factors to the magnitude of the delayed hypersensitivity responses (footpad swelling) to the corresponding antigen in the recipients. Three doses of each of four affinity-purified transfer factor preparations were studied. There were no significant differences in the footpad responses by recipients of either oral or subcutaneous transfer factor. These results support proposals for oral administration of transfer factors in clinical trials.