In vitro studies during long term oral administration of specific transfer factor

153 patients suffering from recurrent pathologies, i.e. viral infections (keratitis, keratouveitis, genital and labial herpes) uveitis, cystitis, and candidiasis were treated with in vitro produced transfer factor (TF) specific for HSV-1/2, CMV and Candida albicans. The cell-mediated immunity of seropositive patients to HSV-1/2 and/or CMV viruses was assessed using the leucocyte migration inhibition test (LMT) and lymphocyte stimulation test (LST) in presence of the corresponding antigens, and the frequency of positive tests before, during and after TF administration was studied. The data were stratified per type of test, antigen and the recipients’ pathology, and statistically evaluated. For the LMT, a total of 960 tests were carried out for each antigen dilution, 3 different antigen dilutions were used per test. 240/960 tests (25.4%) were found positive during non-treatment or treatment with unspecific TF, whereas 147/346 tests (42.5%) were found positive when the antigen corresponding to the specificity of the TF administered to the patient was used (P<0.001). When the data were stratified following pathology, a significant increased incidence of positive tests during specific treatment was also observed (0.0001<P<0.05). In the LST (1174 tests), a significant increase of thymidine uptake was observed in the absence of antigen (control cultures), during treatment with both specific and unspecific TF, but also in the presence of antigen and/or autologous serum during specific TF administration (P<0.0001). TF administration also significantly increased the soluble HLA class I antigens level in 40 patients studied to this effect.     

Efficacy of transfer factor in treating patients with recurrent ocular herpes infections

Recurrent ocular herpes is an insoluble problem for the clinician. As cellular immunity plays an important role in controlling herpes relapses, and other studies have shown the efficacy of HSV-specific transfer factor (TF) for the treatment of herpes patients, an open clinical trial was undertaken in 134 patients (71 keratitis, 29 kerato-uveitis, 34 uveitis) suffering from recurrent ocular herpetic infections. The mean duration of the treatment was 358 days, and the entire follow-up period 189121 before, and 64062 days after TF treatment. The cell-mediated immune response to the viral antigens, evaluated by the lymphocyte stimulation test (LST) and the leucocyte migration test (LMT) (P<0.001), was significantly increased by the TF treatment. The total number of relapses was decreased significantly during/after TF treatment, dropping from 832 before, to 89 after treatment, whereas the cumulative relapse index (RI) dropped, during the same period, from 13.2 to 4.17 (P<0.0001). No side effects were observed. It is concluded that patients with relapsing ocular herpes can benefit from treatment with HSV-specific TF.     

Transfer factor prevents relapses in herpes keratitis patients: A pilot study

Transfer Factor is a dialysable moiety obtained from immune lymphocytes. It has been successfully used for the treatment of several viral infections including labial and genital herpes. In the present study, thirty-three patients with low immune response to HSV antigens and suffering from herpes ocular infections were orally treated with HSV-specific transfer factor (TF). Their relapse index was reduced from 20.1 before treatment to 0.51 after TF administration, with only 6/33 patients relapsing. Although this is not a placebo-controlled-randomized study, the results suggest that TF specific for HSV antigens may be efficacious for preventing relapses of ocular herpes infections as has been the case with genital and labial localisations.Abbreviations CMI Cell-mediated immunity - CMV Cytomegalo-virus - EBV Epstein-Barr virus - HIV Human immunodeficiency virus - HK Herpes keratitis - HSV Herpes simplex virus - IRI Individual relapse index - KU Kerato-uveitis - LMT Leucocyte migration test - LST Lymphocyte stimulation test - MIF Migration inhibition factor - RHK Relapsing herpes keratitis - TF Transfer factor     

Use of transfer factor for the treatment of recurrent non-bacterial female cystitis (NBRC): A preliminary report

Results of conventional treatment of female non-bacterial recurrent cystitis (NBRC) are discouraging. Most patients show an unexpected high incidence of vaginal candidiasis, while their cell mediated immunity to Herpes simplex viruses (HSV) and Candida antigens seems impaired, and it is known that the persistence of mucocutaneous chronic candidiasis is mainly due to a selective defect of CMI to Canadida antigens. Twenty nine women suffering of NBRC, and in whom previous treatment with antibiotics and non-steroid anti-inflammatory drugs was unsuccessful, underwent oral transfer factor (TF) therapy. TF specific to Canadida and/or to HSV was administered bi-weekly for the first 2 weeks, and then once a week for the following 6 months. No side effects were observed during treatment. The total observation period of our cohort was 24379 days with 353 episodes of cystitis recorded and a cumulative relapse index (RI) of 43. The observation period during and after treatment was 13920 days with 108 relapses and a cumulative RI of 23 (P < 0.0001). It, thus, seems that specific TF may be capable of controlling NBRC and alleviate the symptoms.     

Cellular immunity in gnotobiotic primates induced by transfer factor.

Human dialyzable transfer factor (TF) was found capable of inducing in vivo (skin test) and in vitro blastogenesis) cellular immunity in gnotobiotic nonhuman primates. Because the animals were gnotobiotic (germ-free) and had not been skin tested previously, these data support the hypothesis that induction of cell-mediated immunity in recipients of TF does not require a “priming” exposure to specific and/or cross-reacting antigens, and that this induction may be due to an antigen-specific informational effect of TF.In addition, these results support the antigenic specificity of TF, in that recipient primates developed cellular reactivity only against donor “positive” but not against donor “negative” antigens.     

Transfer factor as an adjuvant to non-small cell lung cancer (NSCLC) therapy

The rationale for using transfer factor (TF) in lung cancer patients is that the possibility of improving their cell-mediated immunity to tumour associated antigens (TAA) may improve their survival. From Jan 1984 to Jan 1995, 99 non-small cell lung cancer (NSCLC) resected patients were monthly treated with TF, extracted from the lymphocytes of blood bank donors. In the same period, 257 NSCLC resected patients were considered as non-treated controls. The survival rates of the TF treated group appear significantly improved both for patients in stages 3a and 3b, and patients with histological subtype “large cell carcinoma” (P<0.02). Survival of TF treated patients is also significantly higher (P<0.02) for patients with lymphnode involvement (N2 disease). The results of this study suggest that the administration of TF to NSCLC resected patients may improve survival.     

Transfer factor in vitro abrogation of blocking factors and amplification of immunoreactivity directed to soluble tumor associated antigens in the leukocyte adherence inhibition assay

Summary 26 breast cancer patients with recurrent disease were studied in order to evaluate whether transfer factor (TF) could abrogate serum blocking factors (SBF) and transfer or amplify immune reactivity directed to tumor-associated antigens (TAA) by using the leukocyte adherence inhibition assay (LAI). When 11 leukocyte samples obtained from patients reactive to cancer extract were preincubated with autologous blocking sera in the presence of PBS, nonspecific TF, or specific TF, leukocytes from these patients gained reactivity against breast cancer extract in none out of 11, one out of 11, and nine out of 11 experiments, respectively. Specific TF was obtained from donors with positive LAI breast cancer extract. Mean percentage LAI differences between control extract and breast cancer extract were –4.5±2.6, 0.5±1.7, and 15.5±2.4. The results of specific TF were significantly different from those of nonspecific TF (P<0.005).When nine leukocyte samples obtained from patients unreactive to cancer extract were preincubated with autologous blocking sera in the presence of PBS, nonspecific TF, or specific TF, leukocytes from these patients gained reactivity against breast cancer extract in none out of nine, one out of nine, and seven out of nine experiments, respectively. Mean percentage LAI differences between control extract and breast cancer extract were –4.6±2.0, 0.6±2.3, and 11.8±3.5. The results of specific TF were again significantly different from those of nonspecific TF (P<0.005).When allogeneic blocking sera were utilized, similar results were obtained. When specific TF was administered in two doses, 1 week apart subcutaneously to six breast cancer patients who were unreactive to breast cancer extracts, four patients gained reactivity against breast cancer extracts.Our in vitro experiments support the presence of a specific component in transfer factor extract which can transfer or amplify cell-mediated reactivity with abrogation of SBF directed at TAA.Abbreviations BSS = Balanced Salt Solution - LAI = Leukocyte Adherence Inhibition - PBS = Phosphate Buffered Saline - RPMI = Roswell Park Memorial Institute - SBF = Serum Blocking Factors - TAA = Tumor Associated Antigen - TF = Transfer Factor     

紫外线对白念珠菌的影响

紫外线作为重要的环境因子之一,能显著影响包括白念珠菌在内的多种生物的生长及生理过程.研究发现白念珠菌受紫外线照射后菌丝形成被抑制,孢子形成增多且没有向光性;脉冲紫外线辐射可通过多靶点程序使白念珠菌失活;rad 51缺陷株比rad 52缺陷株更易受紫外线损害,同时紫外线可导致白念珠菌杂合性丢失.研究还发现UVC治疗可明显减少烧伤后真菌微生物感染,核黄素/UVA治疗可明显抑制白念珠菌生长.因此紫外线对白念珠菌有一定的抑制作用.     

Orally administered HSV-specific transfer factor (TF) prevents genital or labial herpes relapses

Forty-four patients suffering from genital (22) and labial (22) herpes were orally treated with HSV-1/2-specific transfer factor(TF). TF was obtained by in vitro replication of a HSV-1/2-specific bovine dialysable lymphocyte extract. Treatment was administered bi-weekly the first 2 weeks, and then weekly for 6 months, most patients received 2–3 courses. The total observation period for all patients before treatment was 26660 days, with 544 relapses, and a relapse index of 61.2, whereas the cumulative observation period during and after treatment was 16945 days, with a total of 121 relapsing episodes and a cumulative RI of 21.4 (P<0.0001). Results were equally significant when the 2 groups of patients (labial and genital) were considered separately. These observations confirm previous results obtained with bovine HSV-specific TF, and warrant further studies to establish HSV-specific TF as a choice of treatment for preventing herpes recurrences.     

Kinetics of Anti-Mannan Antibodies Useful in Confirming Invasive Candidiasis in Immunocompromised Patients

In studying the anti-mannan antibodies longitudinally in serial serum samples of three immunocompromised patients, it was observed that anti-mannan antibodies started to increase shortly after the moment that cultures of deep-tissue sites became positive with Candida albicans. The mean anti-mannan antibody titers determined in a group of 36 immunocompromised patients with invasive candidiasis increased within two weeks after the probable onset of invasive candidiasis. In contrast, anti-mannan antibody levels in serial serum samples of 14 immunocompromised patients who were only colonized with C. albicans remained stable or decreased over time. The HA test measuring the anti-mannan antibodies was 64% sensitive and 89% specific in determining invasive candidiasis. In contrast, antibodies specific for candidal cytoplasmic antigens or enolase alone were of little value in confirming invasive candidiasis in these immunocompromised patients.     

Human T lymphocytes become glucocorticoid-sensitive upon immune activation

A murine model for Transfer Factor (TF) was used in an attempt to identify the nature of its antigen-specific component. TF was prepared from lymph node cells of CBA/Ca/T6 mice sensitized 30 days previously with 2,4-dinitrofluorobenzene (DNFB). To assay for the specific component of TF, 2 × 10⁷ lymphocyte equivalents were injected intravenously into normal syngeneic recipients. Lymph node cells obtained 18–24 hr later gave a positive response in the macrophage migration inhibition (MMI) test in the presence of the soluble analog of DNFB (sodium 2,4-dinitrobenzenesulfonate). The activity of TF was abrogated by absorption with anti-Ia sera including both an Ia alloantiserum (A.TH anti-A.TL) and a xenogeneic rabbit anti-serum which exclusively recognizes carbohydrate-defined Ia antigens. Analysis by paper chromatography using the technique for purification of carbohydrate-defined Ia antigens revealed that MIF production was obtained exclusively with those fractions known to contain Ia antigenic activity. In addition, pretreatment of TF with insoluble conconavalin A (Con A) which has an affinity for carbohydrate-defined Ia antigens resulted in removal of its activity. Taken together these findings pointed to the presence in TF of I-region gene products. Absorption with antibody directed against the dinitrophenyl determinant abolished the capacity of TF to stimulate macrophage inhibition factor production suggesting that it might also contain antigen fragments possibly in association with Ia. No evidence was, however, obtained for H-2 restriction of the action of TF in vivo since it was found to exert an effect in a variety of strain combinations including A.TH and Balb/c which share no known common I-region specificities. Parallel experiments were carried out with the lymphocyte transformation assay since this is known to be a measure of the nonspecific components in TF. Pretreatment with mouse allo-anti-Ia^k serum directed against both protein-and carbohydrate-defined Ia antigens caused a partial reduction in the proliferative response. In contrast no change in response was observed when the TF was absorbed with insoluble Con A or anti-DNP serum. Furthermore, lymphocyte transformation was obtained with only one of the three paper chromatography fractions positive in the MMI assay as well as two other different fractions. Taken together, these findings permitted a distinction to be made between specific and nonspecific components of TF and indicated that the specificity of TF could be explained in terms of the presence of I-region gene coded products possibly in association with antigen fragments.     

Cell-mediated immunity in Cryptococcosis

Cell-mediated immune responses in patients who had recovered from cryptococcosis were compared to those of healthy subjects. Cryptococcal patients were mildly lymphopenic but showed no defect in percentage of thymus-derived lymphocytes. One-third had positive delayed skin test reactions to cryptococcal antigen. Their skin test reactivity to two commonly used noncryptococcal antigens was less intense than healthy control subjects. Strongly positive and specific lymphocyte transformation occurred in the presence of an extract of Cryptococcus neoformans (cryptococcin) in half of the patients. In contrast, few healthy subjects had positive transformation responses to cryptococcin. One patient who was followed sequentially through treatment of cryptococcal meningitis acquired strong cryptococcin reactivity during the course of treatment. Cellular immunologic response to cryptococcin identifies many subjects who have had C. neoformans exposure, and may be of value for assessing immunologic status of patients undergoing therapy. These studies also indicate that most patients with cryptococcosis have a degree of deficiency in cell-mediated response to fungal antigens even when a specific underlying disease process cannot be identified.     

Antisense oligonucleotide for tissue factor inhibits hepatic ischemic reperfusion injury

Tissue factor (TF) is an initiation factor for blood coagulation and its expression is induced on endothelial cells during inflammatory or immune responses. We designed an antisense oligodeoxynucleotide (AS-1/TF) for rat TF and studied its effect on hepatic ischemic reperfusion injury. AS-1/TF was delivered intravenously to Lewis rats. After 10 h, hepatic artery and portal vein were partially clamped. Livers were reperfused after 180 min and harvested. TF expression was studied using immunohistochemical staining. One of 10 rats survived in a 5-day survival rate and TF was strongly stained on endothelial cells in non-treatment group. However, by treatment with AS-1/TF, six of seven survived and TF staining was significantly reduced. Furthermore, we observed that fluorescein-labeled AS-1/TF was absorbed into endothelial cells. These results suggest that AS-1/TF can strongly suppress the expression of TF and thereby inhibit ischemic reperfusion injury to the rat liver.     

Spatial distance and reactivity traits alter the positive perception of brushing by ewes

Gentle handling seems to elicit positive states in sheep. The study investigated whether spatial distance alters sheep responses to brushing and whether spatial distance is influenced by reactivity. Twenty Romane ewes were assessed in three sessions: in Sessions 1 and 3, one grid separated the test animal from pen mates, with no distance between them, and in Session 2 two grids separated the test animal from pen mates by a distance of about 1.7 m. Ewes had been genetically selected for low (R−) or high (R+) behavioural reactivity to social isolation. Body postures, head orientation, ear postures, closed and half-closed eyes, tail wagging and feeding behaviour, in addition to heart rate (HR) and HR variability, as the root mean square of successive differences (RMSSD), standard deviation of all normal-to-normal (NN) intervals (SDNN), RMSSD/SDNN ratio and ratio between low-frequency (LF) and high-frequency (HF) powers (LF/HF) were assessed. Data were analysed using generalized linear models and linear mixed models. Session, genetic line and phase (pre-, brushing and post-brushing) were considered fixed effects. Increased distance in Session 2 might not have influenced ewes’ responses. Fewer changes in ear postures were noted in Session 3 than 1 (P<0.01), suggesting that ewes were more relaxed in Session 3. The RMSSD/SDNN ratio was higher mainly during brushing in Sessions 1 and 3 (P<0.05), indicating that ewes were more relaxed during brushing, and at no distance between pen mates. However, spatial distance influenced R− and R+ ewes’ responses; R+ ewes performed more asymmetric ear postures in Session 2 than 1 and 3 (P<0.01), and in Session 3 than 1 (P<0.01), indicating that spatial distance had a negative effect on R+ ewes. Low reactive ewes spent less time on horizontal ear postures in Session 2 than 1 and 3 (P<0.01), and R+ ewes spent more time on horizontal postures in Session 1 than 3 (P<0.01). Curiously, R− ewes spent more time eating and ruminating in Session 3 than 1 (P<0.01), and in Session 2 than 1 and 3 (P<0.01), whereas R+ ewes ate and ruminated more in Session 1 than 3 (P<0.05). Higher HR was found among R− ewes in Session 2 than 1 and 3, and in Session 3 than 1 (P<0.01). High reactive ewes showed higher HR in Session 1 than 3 (P<0.01). The findings suggest that the social context might influence sheep responses to gentle handling, and the effects depend on their reactivity traits.     

Cytological changes in oral mucosa in denture stomatitis

The cytological patterns associated with the atrophic and hyperplastic forms of denture stomatitis (DS) were studied in 94 patients with DS and 33 controls. Forty percent of patients with DS and 30% of patients in the control group had a positive culture for Candida. When compared to the smears from the control group patients, the smears from patients with DS presented a higher amount of: i) cytological cellular material; ii) fungal cells; iii) cells of the intermediate and parabasal types; iv) cells of the intermediate type with a positive culture for Candida; and v) polymorphonuclear leukocytes, preferentially in association with a positive culture for Candida. Conversely, smears from the control group showed a higher percentage of cells of the superficial type than those of the patients with DS. Although no specific changes in the DS-affected mucosa have been observed by cytology, we consider that this is a useful, easy and inexpensive technique that gives important information about the inflamed mucosa it can be used in the treatment and control of these patients.     

Topical application of a corticosteroid destabilizes the host-parasite relationship in an experimental model of the oral carrier state of Candida albicans

Abstract Using an experimental model in the mouse we have shown that both local and central lines of defense, involving CD4⁺ T cells, participate in a dynamic interaction to maintain a long-term carrier state of Candida albicans in the oral cavity. We have tested the impact of a predisposing factor to oral candidiasis in the form of a topical application of a corticosteroid (Topsyn^® gel) to the oral mucosa for 75 mice twice a day for a 20-day period. Very rapidly after the treatment was initiated, i.e. on day 4, the residual population of Candida increased up to 40-fold and by day 21, the population was 400-fold that of the carrier state. The resident population of intraepithelial CD4⁺ T cells in the oral mucosa virtually disappeared during the treatment. A topical corticosteroid application also resulted in a massive depletion of T cells in the lymph nodes and in the transient abrogation of the DTH reaction to Candida antigens. On cessation of treatment, normal levels of both Candida and intraepithelial CD4⁺ T cells were also quickly restored. These results suggest that resistance to superficial invasion by Candida is linked to the presence of an oral mucosal line of defense and that topical application of corticosteroids may dramatically shift the host-parasite relationship in favor of Candida .     

G试验在侵袭性念珠菌病中的临床价值

目的探讨G试验在侵袭性念珠菌病(invasive candidiasis,IC)中的诊断价值及其与患者粒细胞、病原菌、体温之间的关系。方法选取暨南大学附属五邑中医院2015年9月至2018年6月期间55例IC患者、42例侵袭性曲霉菌病(invasive aspergillosis,IA)患者、30名念珠菌黏膜定植者、50名健康人群,采用免疫比浊法检测各组G试验浓度;采用ROC曲线分析G试验在IC患者中的诊断价值;同时比较G试验阳性率与IC患者粒细胞、病原菌、体温之间的关系;结果采用SPSS 19.0进行统计分析。结果 IC患者G试验水平显著高于健康人群、念珠菌黏膜定植患者和IA患者(P<0.05)。55例IC患者中,非白念珠菌例数(32例)超过白念珠菌(23例)。分别以非侵袭性真菌组(invasive fungal disease, IFD)和非IC组作为对照组,G试验诊断IC患者的ROC曲线下面积(AUC)为0.899和0.782。粒细胞正常组IC患者G试验阳性率与粒细胞增高组、粒细胞减少组之间无统计学差异(P>0.05),但显著高于粒细胞缺乏组(P<0.05);白念珠菌IC组G试验阳性率与热带和光滑念珠菌组差异无统计学意义(P>0.05),但显著高于其他念珠菌感染组(P<0.05);G试验阳性率在不同体温组IC患者均无统计学差异(P>0.05)。结论 G试验在IC患者中具有较高的诊断价值,不仅可区分致病菌和定植菌,还对IC和IA的鉴别具有一定的价值;G试验阳性率与IC患者的粒细胞缺乏和感染的病原菌类型相关,与体温无关。     

Immunogenicity of synthetic TF-KLH (keyhole limpet hemocyanin) and sTn-KLH conjugates in colorectal carcinoma patients

Mucins of colorectal carcinomas overexpress the cancer-associated disaccharides Thomsen-Friedenreich antigen (TF) and sialyl-Tn antigen (sTn), making these antigens suitable for active specific immunotherapy. Patients at high risk for recurrent colon cancer, but free from disease after surgical resection, were immunized with synthetic TF and sTn covalently attached by a two-carbon crotyl linker to keyhole limpet hemocyanin (KLH). Four groups of patients were treated with TF-KLH without adjuvant, TF-KLH plus the immunological adjuvant Detox, sTn-KLH plus Detox, or sTn-KLH plus the immunological adjuvant QS-21, and the serological response was monitored. Enzyme-linked immunosorbent assay (ELISA), do-blot immunostains, and inhibition assays were used to identify antibody responses against synthetic TF and sTn epitopes and against natural antigens, including asialoglycophorin expressing TF antigen, and ovine submaxillary mucin and the human colon cancer line LS-C expressing sTn antigen. Our results demonstrate that vaccines containing TF or sTn-KLH conjugates plus immunological adjuvants Detox and especially QS-21 induced high IgM and IgG antibody titers against the respective synthetic disaccharide epitopes. However, when tested against natural antigens expressing these disaccharide epitopes, IgM antibodies showed weak to moderate reactivity, while IgG antibodies were almost totally unreactive. On the basis of these results we are continuing to test modifications of synthetic TF and sTn epitopes to identify those that induce IgM and IgG antibodies that are more reactive with these antigens as they are expressed on tumor mucins.This work was supported by grants from the National Institutes of Health (CA33049, CA52491, CA08748) and the Chemotherapy Foundation     

In Vivo Transfer of cellular immunity to primates with transfer factor prepared from human or primate leucocytes

Dialyzable transfer factor (TF_d) prepared from a human donor and transfer factor (TF) from baboon whole cell lysates was administered to 3 species of nonhuman primates: baboons, cebus monkeys and marmosets. In vivo transfer was evaluated with in vivo skin test and in vitro blastogenic responses to multiple antigens. Transfer of cellular reactivity in all three nonhuman primate species was demonstrated with both human TF_d and baboon TF. A cumulative conversion rate of 45% for skin test responses and 65% for lymphocyte blastogenesis was demonstrated following human TF_d injection while conversion was 17% and 33% respectively following baboon TF. Specificity was supported by the absence of conversion to TF donor negative antigens. There were no signficant differences observed between the 3 recipient primate species.     

Transfer factor with anti-EBV activity as an adjuvant therapy for nasopharyngeal carcinoma: A pilot study

Overall survival of nasopharyngeal carcinoma (NPC) at UICC stage IV still remains unsatisfactory even with combination chemotherapy (CT) and radio-therapy (RT). In view of the association of reactivation of Epstein-Barr virus (EBV) with the development and recurrence of NPC, immunotherapy in the form of transfer factor (TF) with specific activity against EBV (TF-B1) was suggested as an adjuvant to a combination of CT and RT in order to improve survival. In the present study, 6 UICC Stage IV patients received TF-B1 and another 6 patients matched for disease stage were given TF prepared from peripheral blood leucocytes (TF-PBL). Results were compared with another 18 patients matched by age, sex, and stage of disease who received standard therapy without TF during the same period (C group). After a median follow up of 47.5 months, the survival for the TF-B1 group was found to be significantly better (P=<0.05) than the PBL and C group. While the 8 patients with distant metastasis (DM), not treated with TF-B1 (6 in the control and 2 in the PBL group), died due to progressive disease (average survival being 14.3 months), both patients with DM in the TF-B1 group had complete remission: one died of tuberculosis after surviving for 3.5 years and another is still alive, disease free, after 4.2 years. Although the series involved a small number of cases, the apparent effect of adjuvant immunotherapy in the form of TF with anti-EBV activity is of considerable interest.