Biologic and molecular characterization of the IgG serum blocking factor (SBF-IgG) isolated from sera of patients with EBV-induced infectious mononucleosis

Our laboratory has previously reported the isolation of a serum blocking factor (SBF) from infectious mononucleosis (IM) patients. The SBF has been purified by a combination of Sephadex QAE-50 ion exchange and Sephadex G-200 molecular sieve chromatography. This material was found to be devoid of soluble immune complexes, and immunochemically and biochemically was characterized as IgG and, hence, termed SBF-IgG. The SBF-IgG was shown to significantly (alpha = 0.05) suppress antigen specific (Influenza A-1[H1N1]) in vitro lymphocyte stimulation (LS) as well as leukocyte migration-inhibition (LMI) reactivity. Also, the SBF-IgG significantly suppressed the in vitro.LS response to phytohemagglutinin. In addition, the SBF-IgG when bound to normal donor lymphocytes significantly reduced the high affinity E-rosette (HAR) reactivity at 29 degrees C. A purified T lymphocyte subpopulation of normal donor lymphocytes specifically bound SBF-IgG, and the latter could be r covered using glycine-HCI. It appears that SBF-IgG is a nonspecific antibody; it binds neither lymphokines nor specific antigen, but apparently elicits its in vitro vitro cell-mediated suppressive effect at the level of the T lymphocytes.     

A preliminary report on the use of transfer factor for treating stage D3 hormone-unresponsive metastatic prostate cancer

As conventional treatments are unsuccessful, the survival rate of stage D3 prostate cancer patients is poor. Reports have suggested the existence of humoral and cell-mediated immunity (CMI) against prostate cancer tumour-associated antigens (TAA). These observations prompted us to treat stage D3 prostate cancer patients with an in vitro produced transfer factor (TF) able to transfer, in vitro and in vivo, CMI against bladder and prostate TAA. Fifty patients entered this study and received one intramuscular injection of 2–5 units of specific TF monthly. Follow-up, ranging from 1 to 9 years, showed that complete remission was achieved in 2 patients, partial remission in 6, and no progression of metastatic disease in 14. The median survival was 126 weeks, higher than the survival rates reported in the literature for patients of the same stage.     

Tartrate-resistant acid phosphatase 5b as a serum marker of bone metastasis in breast cancer patients

Diagnosis and follow-up of bone metastases in breast cancer patients usually rely on symptoms and imaging studies. Tartrate-resistant acid phosphatase 5b (TRACP 5b) is a specific marker of osteoclasts and is herein proposed as a marker of bone metastasis in breast cancer patients. An immunoassay using a monoclonal antibody, 14G6, was used to measure the activity of serum TRACP 5b at pH 6.1 in 30 early breast cancer patients without bone metastasis and in 30 aged-matched breast cancer patients with bone metastasis. Another 60 normal volunteers were recruited as controls. Bone alkaline phosphatase (BAP), a traditional marker of bone turnover, was also measured in selected cases. The overall mean TRACP 5b activity in normal women was 2.83 ± 1.1 U/I, and it increased with age. The mean TRACP 5b activity in early breast cancer patients did not differ from that of the normal group (2.93 ± 0.64 vs. 2.83 ± 1.1 U/I; p=0.66), whereas it was significantly higher in breast cancer patients with bone metastasis (5.42 ± 2.5 vs. 2.83 ± 1.1 U/I; p<0.0001). BAP activity was significantly higher in breast cancer patients with bone metastasis than in early breast cancer patients (p=0.004). Serum TRACP 5b activity correlated well with BAP activity in breast cancer patients with bone metastasis (p<0.0001), but not in normal individuals or in patients without bone metastasis. TRACP 5b activity can be considered a surrogate indicator of bone metastasis in breast cancer patients.     

Conversion of human lymphocytes to antitumor immune reactivity by xenogeneic and allogeneic imune RNA detected by leukocyte-adherence inhibition tests

Summary Using leukocyte adherence inhibition (LAI) tests, we studied the activity of xenogeneic immune RNA (I-RNA) extracted from the spleen and lymph nodes of sheep after immunization with human breast carcinoma tissue or keyhole limpet hemocyanin (KLH) in inducing lymphocytes from normal healthy donors to mediate immune responses in vitro. Mononuclear cells isolated from venous blood of normal donors, depleted of monocytes and, in some experiments, separated into T cells and non-T cells, were incubated with and without anti-breast carcinoma I-RNA or anti-KLH I-RNA for 20 min at 37° C. Then, lymphocyte adherence was determined by a Coulter counter method in the presence of 3 M KCl extracts of breast carcinoma tissues, control tissue, or KLH. Following incubation with anti-breast carcinoma I-RNA, the adherence of lymphocytes from normal donors was found to be inhibited only in the presence of breast carcinoma extracts. Following incubation with anti-KLH I-RNA, lymphocyte adherence was inhibited only in the presence of KLH. The principal effector cells involved appeared to be T lymphocytes. I-RNA treatment with RNase, but not with DNase or pronase, completely abrogated the LAI responses. In a blind study utilizing coded samples of xenogeneic and allogeneic I-RNA of unknown origin, samples containing activity against breast cancer extracts were identified correctly by LAI. Abbreviations used: I-RNA, immune RNA; LAI, leukocyte adherence inhibition; KLH, Keyhole limpet hemocyanin; PBS, phosphate buffered saline; RNase, ribonuclease; DNase, deoxyribonuclease     

Expression by human fetal organs of organ-specific cancer neoantigens as measured by leukocyte adherence inhibition

Summary Human cancers express organ-specific cancer neoantigens (OSN) as determined by in vitro leukocyte responses to extracts of cancers by the tumor host. In this study, we determined whether the OSNs were normal developmental proteins that were expressed by fetal organs and re-expressed with oncogenesis. Fetal extracts, principally of lung and colon but also of liver and kidney, were tested for their ability to induce leukocyte adherence inhibition (LAI) as compared to extracts from adult tissues of the same organ. Leukocytes from lung cancer patients showed positive LAI responses to 13- and 19-week fetal lung tissue. Likewise, leukocytes from colon cancer patients showed positive LAI responses to 14- and 19-week fetal colon tissue, whereas leukocytes from control subjects did not. Neither group responded positively to 21-week fetal organs. Criss-cross experiments showed that the fetal antigen was organ specific. Multiparous pregnant women showed positive LAI responses to cancer extracts but not to extracts from normal tissues of the same organ. The pattern of the LAI response was bell-shaped. Positive LAI responses to lung and breast cancer were detected at 4 to 7 months gestation and peaked at 5 months. To the fetal colon, LAI positive responses were detected at 5 to 8 months gestation, with the peak response at 6 months. The results indicate that OSN of cancers are also expressed by fetal organs and sufficient antigen is shed by fetal organs to sensitize pregnant women. Older fetal organs (21 weeks) and adult organs do not express an immunogenic or antigenic OSN.Supported by a grant from the National Cancer Institute of Canada     

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.     

Cross-reactivity of lung cancer patients' leucocytes in the leucocyte migration inhibition test

Summary Panels of 3 M KCl extracts of squamous-cell carcinomas, adenocarcinomas and oat-cell carcinomas of the lung were used for a comprehensive analysis of cross-reactivity in the leucocyte migration test. Lung cancer patients' leucocytes showed positive reactivity in 69%–100% of cases (n=353). No significant differences were observed when data were grouped with respect to the histological type of the tumours used for extraction or of the tumours of the leukocyte donors. Leukocytes of patients bearing tumours of nonpulmonary origin exposed to lung cancer extract panels and leukocytes of lung cancer patients exposed to gastrointestinal cancer extract panels were definitely less reactive (35%–47% and 6%–38%, respectively). However, a high reaction frequency was found in patients with lung metastases from different nonpulmonary tumours. This group of patients also frequently showed reactivity (52%) with normal lung tissue extracts. Patients with benign lung diseases reacted positively with lung tumour extracts in 25%–39% of cases, but donors with other benign disease and healthy controls were virtually nonreactive (0–14%).Hence, a high degree of cross-reactivity occurs in the lung cancer system and restricted cross-reactivity occurs with tumours of other organs. Possible explanations for the lung-oriented reactivity of patients with lung metastases are discussed.Abbreviations LMI leucocyte migration inhibition - MI migration index - LMT leucocyte migration test - SCC squamous-cell carcinoma - OCC oat-cell carcinoma - AC adenocarcinoma     

Amplification of lymph node cell tube leukocyte adherence inhibition (LAI) reactivity by leukocyte adherence inhibition factor (LAIF)

This investigation examines the immunologic basis for specific antigen-induced tube leukocyte adherence inhibition (LAI) reactivity of draining lymph node cells (LNC) from dogs with canine transmissible venereal sarcoma (CTVS). CTVS regressor LNC, macrophage-depleted LNC, and enriched T lymphocyte fractions, but not enriched B lymphocyte fractions, were specifically reactive to CTVS antigen extract in direct tube LAI. In addition, regressor LNC amplified tube LAI responses by generating supernatants with leukocyte adherence inhibition factor (LAIF) activity for normal dog indicator LNC and enriched peripheral blood mononuclear cells (PBMC) in an indirect tube LAI assay. However, macrophage-depleted LNC and enriched T lymphocyte fractions failed to generate supernatants with LAIF activity, suggesting that macrophage accessory cells play a central role in the amplification of tube LAI. Interestingly, CTVS regressor peripheral blood leukocytes (PBL) and PBMC, which were specifically reactive in direct tube LAI, also failed to generate supernatants with LAIF activity. These findings demonstrate a distinction between LAIF-mediated amplification and direct tube LAI reactivity, and suggest that leukocyte populations with differing cellular proportions and from different immunologic compartments may participate in tube LAI via different mechanisms.     

Transmembrane signal defect and absence of cancer extract induced leukocyte adherence inhibition (LAI) for leukocytes from patients with advanced cancer

Summary Leukocytes from patients with early cancer exhibit leukocyte adherence inhibition (LAI) when incubated with extracts of cancer of the same organ and histogenesis, whereas leukocytes from patients with advanced cancer seldom do. To understand the reason for this refractory state, tumor antigen-induced LAI and transmembrane signalling were measured in the same leukocytes. Transmembrane signalling was measured by changes in membrane potential () by the [³H]tetraphenylphosphonium equilibration technique. When leukocytes from patients with early breast cancer were incubated with extracts of breast cancer and malignant melanoma they showed changes consisting of depolarization and hyperpolarization beginning within 0.5 min after addition of the breast cancer extract and finishing 15 min later. Moreover, they showed no changes when incubated with extracts of normal breast tissue. Leukocytes from subjects without cancer seldom showed changes. In criss-cross experiments, leukocytes from patients with melanoma only exhibited changes when incubated with the melanoma extract. There was a strong correlation between cancer extract-induced change and LAI. The change was triggered by leukotriene-like mediators from antibody-dependent monocytes. Authentic leukotrienes triggered changes in all subpopulation of leukocytes. Leukocytes from patients with advanced breast cancer when incubated with breast cancer extract did not transmit a signal or show LAI. Brief elevation of intracellular cyclic AMP restored both change and LAI induced by breast cancer extracts, indicating that reactive leukocytes are present but in a refractory state. We conclude that leukocytes from patients with advanced cancer do not react in LAI because tumor antigen does not trigger a transmembrane signal to initiate the cascade of biochemical reactions and physiological changes for LAI.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - cyclic AMP cyclic adenosine monophosphate - ETYA eicosatetraynoic acid - HEPES 4-(2-hydroxyethyl)-1-piperazine ethansulfonate - LAI leukocyte adherence inhibition - NAI nonadherence index - OSN organ-specific cancer neoantigen - PBL peripheral blood leukocytes - PGE₂ prostaglandin E₂ - [³H]TPP⁺ [phenyl³H]tetraphenylphosphonium bromide - transmembrane potential     

Leukocyte migration inhibition test in breast cancer patients treated with conventional therapy or with conventional therapy plus poly A · poly U

Summary Leukocyte migration inhibition tests (LMIT) were performed to measure the immunologic reactivity of patients with initially operable breast cancer. Leukocyte migration was inhibited in 38% of 159 patients in the presence of autologous tumor extract, in 33% of 160 subjects in the presence of autologous serum, and in 47% of 148 patients in the presence of extract and serum. These patients formed part of a randomized clinical trial comparing conventional treatment complemented by injections of poly A · poly U with conventional treatment and injection of saline as a placebo.A sequential study was carried out, testing the leukocyte reactivity 7 days, 2 months, 4 months, and 1 year after the operation. The percentage of patients with positive LMIT increased significantly with time. Statistical comparison revealed no significant difference in the reaction of the two therapeutic groups. In addition, no significant difference was found between those with lymph node involvement and those without, although the percentage of positive LMIT in the presence of tumor extract appeared to be higher in the second group 1 year after surgery. Radiotherapy given to those with lymph node involvement did not significantly change their reaction.This study also confirmed the presence in some autologous sera of a synergistic factor (SS factor) that increased the inhibition of migration of leukocytes by autologous tumor extract. This factor was found in 18 patients, who were equally distributed between the two therapeutic groups.In the group with synergistic factor, the percentage with lymph node involvement appeared greater (83% compared with 68% among those patients who had no SS factor), and the incidence of distant metastases was also raised (44% compared with 21%). This factor seemed to indicate a poor prognosis. Moreover, there was a difference in the results between the two therapeutic groups in patients with synergistic factor. Of nine patients undergoing conventional treatment, six developed metastases, whereas only two of the nine patients also receiving poly A · poly U developed metastases. The same trend was observed throughout the trial population.     

组织因子阳性的细胞和微粒在胃癌高凝状态中的作用

目的:探讨胃癌患者血浆中组织因子阳性的血小板、白细胞和微粒的数量及其促凝活性。方法:将45例胃癌患者根据TNM分期分为Ⅰ、Ⅱ、Ⅲ、Ⅳ期,同时选取30例健康人作为对照组。采用流式细胞术检测组织因子阳性的细胞和微粒数。凝血酶生成实验检测细胞和微粒的凝血活性。结果:胃癌Ⅲ/Ⅳ期患者血浆中组织因子阳性的血小板、中性粒细胞、单核细胞和微粒的数量明显高于胃癌Ⅰ/Ⅱ期和健康对照组。胃癌Ⅲ/Ⅳ期患者血小板、白细胞和微粒的促凝活性与其他组相比显著升高,与增加凝血酶的生成速度和生成总量有关。用抗组织因子抗体抑制TF后,细胞和微粒的凝血活性明显下降。然而,使用抗膜连蛋白V抑制PS后,细胞和微粒的凝血活性虽然有下降趋势,但是并不明显。此外,根治性手术治疗可以降低组织因子阳性的血小板、中性粒细胞、单核细胞和微粒的数量。结论:组织阳性的血小板、中性粒细胞、单核细胞和微粒是胃癌Ⅲ/Ⅳ患者高凝状态的原因之一,通过抑制TF和凝血酶的生成可能降低胃癌患者的血栓发生率。     

Regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with progressing or spontaneously regressing canine transmissible venereal sarcoma (CTVS)

Summary We examined the regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with regressing or progressing canine transmissible venereal sarcomas (CTVS). Both regressor and progressor peripheral blood leukocytes (PBL), draining and nondraining lymph node cells (LNC), and splenic leukocytes were significantly responsive to CTVS antigen extract in tube LAI. In contrast, a significant decrease in basal glass adherence of progressor PBL, draining and nondraining LNC, and splenic leukocytes was observed. Normal glass adherence was restored to progressor leukocytes by extensive washing with warm serum-free media, while significant tube LAI responsiveness to CTVS antigen extract was maintained. Preincubation of regressor PBL and LNC with progressor sera in two-stage tube LAI decreased the basal glass adherence of treated leukocytes. This effect of progressor sera was heat labile, a characteristic of CTVS antigen. Collectively, these findings suggest that progessor leukocytes and progressor sera treated regressor leukocytes were activated by interaction with serum CTVS antigen and thus behaved in tube LAI as stimulated cells, even in the absence of CTVS antigen. Regressor but not progressor sera were shown to contain anti-CTVS IgG with specific arming activity for normal dog PBL, but not LNC in two-stage tube LAI. The nonadherent response of peripheral blood neutrophils in two-stage tube LAI was proportional to the concentration of arming IgG, whereas no change was observed in glass adherence of PBL. The results of this study define the role of progressor and regressor serum factors in the mechanism of tube LAI and demonstrate a relationship between leukocyte glass adherence and the clinical course of CTVS. These findings show that tube LAI is a simple and reproducible measure of active factors in the immune response to a tumor.This investigation was supported in part by grant, CA-23469, from the National Cancer Institute, DHHS, and is submitted as Scientific Contribution No. 1051, Storrs Agricultural Experiment Station, University of Connecticut, Storrs, CT 06268 USA     

Adherent indomethacin-sensitive suppressor cells in malignant melanoma

Summary Twenty-five patients with malignant melanoma were studied to determine the mechanisms underlying decreased cellular immunity in this disease. Sixteen patients were examined following surgical resections of tumor, and nine patients had evidence of metastases. Patients with metastatic disease (group II) had decreased lymphocyte transformation to concanavalin A (48,255±30,074) compared with controls (83,550±41,277) and patients free of disease (group I) 110,231±59,990) (P>0.01). Rigorous depletion of monocytes by adherent techniques resulted in an improvement in blastogenesis in seven of nine group II patients, whereas controls and group I patients had marked decreases in reactivity. Indomethacin also improved lymphocyte reactivity; a mean of 166.4% was recorded in five metastatic disease patients studied, as against a mean of 2% in group I and 9.65% in controls. In seven patients followed serially the presence of adherent suppressor cells tended to correlate with progressive disease, shorter survival, and decreased skin test reactivity.     

Evaluation of serum lipids and high-density lipoprotein subfractions (HDL2, HDL3) in postmenopausal patients with breast cancer

Breast cancer patients are known to be at increased risk for developing other chronic diseases including cardiovascular disease. Studies by different investigators have shown a correlation between increased dietary fat or hypercholesterolemia and the occurrence of breast cancer. Since previous studies on lipoprotein subfractions in this type of cancer have been inconsistent, we evaluated the lipids and lipoprotein subfraction levels in postmenopausal patients with breast cancer in an attempt to identify the risk for the development of cardiovascular disease.The study included 132 patients, 56 of which were suffering from breast cancer, 32 from pancreatic and 44 age-matched controls. Total cholesterol (TC), triglycerides and lipoprotein fractions as well as TC/High density lipoprotein (HDL) and HDL2/HDL3 ratios were estimated by standard laboratory techniques.An increase in triglycerides and a decrease in HDL-cholesterol, especially in the HDL2 subfraction, were observed in patients with breast cancer as compared to the controls (P < 0.05). The maximum changes in TC, and HDL concentrations were observed in patients with advanced disease. Analysis of indexes of atherosclerosis (TC/HDL, and HDL2/HDL3 ratios) demonstrated that breast cancer patients had significantly higher TC/HDL ratio (6.44 ± 1.24) compared with controls (3.43 ± 0.57, p = 0.001), and patients with pancreatic cancer (3.79 ± 0.15, p = 0.027).The results have demonstrated an unfavourable lipid profile in untreated breast cancer patients with high atherosclerosis indexes. This observation is of great importance, considering the potential use of endocrine therapy that could result in further deterioration of lipid indexes. We propose the evaluation and monitoring of lipid profile prior and after the induction of hormonal therapy in breast cancer patients, as a routine in clinical setting. (Mol Cell Biochem 268: 19–24, 2005)     

Evaluation of human LOX-12 as a serum marker for breast cancer

The high concentration of prostaglandins has been associated with chronic inflammatory diseases and several types of human cancers. This is due to the over expression of inflammatory enzymes like Cyclooxygenase (COX), Lipoxygenase (LOX) etc. The aim of this study was to quantify the LOX-12 with clinicopathological parameter of breast cancer patients and its response after chemotherapy to establish serum LOX-12 as a prognostic marker. This case-controlled study was performed on 86 biopsy proven breast cancer patients. Blood and tissue samples were collected from the patients. Serum LOX-12 of the study group was quantified by Surface Plasmon Resonance (SPR) and ELISA techniques by antibody–antigen interaction strategy. A significant increase in LOX-12 levels was observed in breast cancer patients (Mean ± SD = 40.54 ± 13.61 ng/ml) as compared to healthy controls (Mean ± SD = 13.42 ± 2.4 ng/ml) (p < 0.0001). Serum LOX-12 levels were significantly higher (p < 0.002) in patients with lymph node involvement. More than 75% patients had shown significant (p < 0.0001) reduction of LOX-12 levels after chemotherapy. This was also confirmed by ELISA. This study for the first time had co-related the quantity of serum LOX-12 with breast cancer and also with the effect of chemotherapy.     

Transfer factor and cellular reactivity to varicella-zoster antigen in childhood leukemia

The present studies were designed to evaluate possible efficacy of specific human transfer factor (TF) in preventing or attenuating varicella-zoster (VZ) infection in children with acute lymphocytic leukemia (ALL). TF was prepared following leukapheresis of adult donors who were convalescing from chicken pox. Recipients were VZ seronegative children with ALL, 12 in remission and 3 in relapse; a single injection of TF was given equivalent to 10⁸ lymphocytes per 7 kg body wt. The following VZ-specific parameters were measured: lymphocyte blastogenesis, cytotoxicity and leukocyte inhibitory factor (LIF) production, and indirect fluorescent and CF antibody titers. No patients in relapse converted immune response while $\text{10}\text{12}$ in remission developed positive reactivity in at least one assay of cell-mediated immunity (CMI); $\text{3}\text{12}$ were positive in all three parameters of CMI and $\text{8}\text{12}$ in two assays. Cytotoxicity was the most consistently positive test following TF administration. No patients developed VZ antibody. Seventeen months follow-up has demonstrated persistent cellular immunity.     

Post term dietary-induced changes in DHA and AA status relate to gains in weight, length, and head circumference in preterm infants

Preterms need supplementation with docosahexaenoic (DHA) and arachidonic (AA) acids to prevent steep postnatal declines.Associations between growth and erythrocyte (RBC) DHA and AA were studied in 139 preterms (51% male, gestational age 30.3±1.5 weeks, birth weight 1341±288 g) fed human milk with breast milk fortifier or preterm formula until term, followed by postdischarge formula (PDF; n=52, 0.4% DHA, 0.4% AA), term formula (TF; n=41, 0.2% DHA, 0.2% AA), or human milk (HM; n=46).At six months, PDF resulted in higher RBC-DHA than TF and HM, while RBC-AA was higher than TF, but similar to HM. There were no between-group differences in growth between term and six months. RHC-DHA related positively with gain in weight and length and negatively with gain in head circumference. RBC-AA related positively with gain in head circumference and negatively with gain in weight and length.In conclusion, PDF with higher DHA and AA than TF may promote postnatal growth of preterms.     

Effects of distant metastasis and peripheral CA 15-3 on the induction of spontaneous T cell responses in breast cancer patients

Tumor-specific memory T cells are detectable in the bone marrow (BM) of a majority of breast cancer patients. In vitro they can be reactivated to IFN-γ producing, cytotoxic effector cells and reject autologous, xenotransplanted tumors in NOD/SCID mice after specific restimulation with autologous dendritic cells (DC). In this study, we demonstrate the presence of specific tumor-reactive BM memory T cells in altogether 56 out of 129 primarily operated breast cancer patients by short-term IFN-γ EliSpot assays with unstimulated T cells and tumor antigen presenting, autologous DCs. We observed tumor-reactive BM memory T cells predominantly in patients with primarily metastatic disease (P = 0.011) or with increased concentrations of tumor marker CA 15-3 in the peripheral blood (P = 0.004), respectively. Memory T cell reactivity against HLA-A^*0201-restricted peptides from the tumor-associated antigens MUC1, Hpa_16–24 and Hpa_183–191 was also detected particularly in patients with elevated peripheral CA 15-3 concentrations (P < 0.05). Altogether these data indicate that the systemic presence of tumor-derived antigens promotes an induction of tumor-specific cellular immune responses in the human BM.     

Bone metastasis as a prognostic factor in breast cancer patients with liver metastasis given OK-432-combined adoptive immunotherapy via the hepatic artery

The outcome in 31 patients with liver metastases from breast cancer given OK-432-combined adoptive immunotherapy via the hepatic artery was analyzed. Patients received intraarterial OK-432, a streptococcal preparation, followed by the transfer of autologous lymphocytes cultured with autologous tumor extract and interleukin-2 for 9–13 days. Liver lesions were evaluable in 11 of the 12 patients with bone metastasis (group A) and in 16 of the 19 patients without bone metastasis (group B). Complete response (CR) in the liver was attained in 8 patients in group A, but in only 1 in group B (p < 0.01). In group A, radiological features of all metastatic foci of bone improved after CR in the liver. Moreover, the median survival time (MST) of group A (20 months) was longer (p=0.06) than that of group B patients with extra-hepatic metastasis (n=12; MST=6 months), while group B patients with liver metastasis alone (n=7) showed a MST similar to that of group A. Thus, loco-regional immunotherapy via the hepatic artery was found to be useful in controlling both liver and bone metastasis from breast cancer. Moreover, in breast cancer patients with liver metastasis, bone metastasis appears to be a prognostic factor associated with good response to this immunotherapy.Abbreviations MST median survival time - CR complete response - PR partial response - MDP metyl-diphosphonate     

Soluble cell membrane antigens associated with bladder cancer

Summary Separated soluble membrane components present on bladder cancer cells were screened for their ability to produce cell-mediated immune responses, and those antigens that were tumor-associated (TAA) were purified and identified. A total of 812 tests of control and cancer antigens were performed in 110 patients. In a series of 384 skin tests performed in 28 patients with crude antigens separated by gel filtration and a series of 322 skin tests performed in 60 patients with semipurified antigens further separated by gradient polyacrylamide gel electrophoresis, the average 48-h induration response in those patients who reacted was not significantly different in relation to the stage of cancer. Preoperative patients responded to a tumor-associated antigen, and postoperative patients responded not only to the tumor-associated antigen, but also to a tissue-associated antigen, which may possibly contain tumor-related components. Of these bladder cancer patients, 78% had a positive delayed hypersensitivity reaction to skin tests with 30 g semipurified bladder cancer TAA, whereas only 53% had reactions to one or more of the recall antigens used, which included SKSD, candida, dermatophytin, PPD, and mumps. TAA was also isolated and identified on bladder cancer tissue culture line T-24. More highly purified bladder TAA was highly specific in controlled skin tests for delayed hypersensitivity reaction to 5 g per test in 20 patients. The amount of TAA present on primary bladder tumor cells is approximately 0.2 pg, and on T-24 cell it is approximately 0.04 pg; this is approximately 2% of the soluble protein on a primary bladder cancer cell and about 0.8% of the soluble protein on a T-24 cell. Reactions with TAA in leukocyte migration inhibition tests were partial; TAA is a very weak reactant in double diffusion tests; TAA shows promise in indirect immunofluorescent tests, in some complement fixation tests, and for use in enzyme-linked immunoadsorbent assays. Bladder cancer TAA is a simple polypeptide, fairly stable, with an estimated molecular size of approximately 40,000 daltons. The tissue-associated antigen reacts in double diffusion with sera from patients with benign and malignant bladder conditions, and is a polypeptide of approximately 80,000 daltons; whether this less specific antigen is a dimer containing the TAA component must still be determined.