Immune competence in breast cancer — Relationship of pretreatment immunologic tests to diagnosis and tumor stage

Summary The immune competence of a group of 276 patients with suspected breast cancer has been assessed using a spectrum of tests: the peripheral lymphocyte count, serum immunoglobulin levels, lymphocyte response to phytohemagglutinin (PHA), Mantoux test, and dinitrochlorobenzene (DNCB) skin test. All tests were completed prior to any form of treatment as the initial part of an ongoing, long-term assessment which will ultimately relate immune competence to prognosis. 225 patients with breast cancer were allocated into stages based on their TNM status. The remaining 51 patients proved to have benign breast disease and made up the control group. In analysis, control patients were compared with early breast cancer patients, while the effect of advancing disease was assessed by betweenstage comparisons in the cancer group.There were no significant differences between early breast cancer and control patients or between cancer stages in peripheral lymphocyte count, serum immunoglobulin levels, lymphocyte response to PHA, or Mantoux responses. Age was found to have a crucial effect on some of these parameters and some apparent differences between the various groups lost significance after appropriate allowances were made for age.Important differences seen with the DNCB test persisted after allowing for age effects. Responses to DNCB were significantly depressed in patients with early breast cancer compared to controls. Patients with disseminated cancer showed greater depression than early breast cancer patients, but surprisingly, patients with locally advanced tumors had good responses to DNCB. Possible reasons for the paradoxical preservation of DNCB reactivity in patients with locally advanced cancer are discussed.The DNCB test is the most discriminating of the five tests of immune function studied.     

Clinical studies on cell-mediated immunity in gestational trophoblastic disease: nonspecific cellular immunocompetence in patients with hydatid mole and trophoblastic neoplasia

The nonspecific cell-mediated immunocompetence of 51 patients with gestational trophoblastic disease (GTD), including 16 patients with hydatid mole (HM), 24 with nonmetastatic trophoblastic neoplasia (NTN) and 15 with metastatic TN (MTN), was studied with the use of both in vitro and in vivo parameters of cell-mediated immunity (CMI) such as lymphocyte blastogenic response to phytohemagglutinin (PHA), subpopulation constitution, and delayed cutaneous hypersensitivity responses to 2,4-dinitrochlorobenzene (DNCB) and to purified protein derivative of tuberculin (PPD). The pretreatment cell-mediated immune status of patients with HM developing no malignant sequelae was shown to be essentially similar, in terms of both in vitro PHA and in vivo DNCB reactivities, to that of normal women and of patients with benign gynecological diseases. In patients with TN, however, there was a significant depression in the blastogenic lymphocyte response to PHA before evacuation of the mole, which was persistently demonstrated after uterine evacuation and more marked throughout the course of disease in patients with MTN, than in those with NTN, with a tendency to return to normal in remission. Moreover, patients with TN had a significant depletion of T lymphocytes as determined by rosette-forming cell procedures before treatment, which was most evident in patients with MTN. Plasma from the MTN patients was also shown to have an inhibitory effect on PHA responsiveness of lymphocytes from normal women. There was an increased incidence of impaired reactivity to DNCB in patients with TN (higher in MTN than in NTN), compared with HM and benign diseases, while no such difference in incidence was observed in response to PPD. On the basis of these findings, a preliminary characterization of altered immunocompetence in patients with TN and its mechanism are discussed.     

Immune functions and the prognosis of patients with solid tumours

Summary The immune competence of 169 patients with solid malignant tumours was assessed before initiation of radiotherapy or chemotherapy and followed during the course of the disease. The data of years 1974–1984 were collected and subjected to an analysis in order to evaluate their prognostic significance. The number of leucocytes and lymphocytes in the peripheral blood, the percentage or absolute number of E-rosette forming cells or EAC-rosette forming cells or serum immunoglobulin levels did not show any association with the prognosis. Lymphocyte proliferative responses to PHA, Con A and PPD as studied before initiation of the treatment did not correlate with recurrence or final prognosis of the disease, except that the responses to PPD were slightly lower in patients with recurrence of gynaecological cancer, melanoma or gastrointestinal cancer than in their respective control patients. In the values observed after the first treatment course a low response to PPD was associated with poor prognosis in patients with melanoma or gastrointestinal cancer. At the time of recurrent disease the PPD response showed an association with a poor final outcome in patients with gastrointestinal malignancy. Of the responses assessed less than 3 months before death due to cancer, only in patients with breast cancer were low Con A responses seen; in all patient groups the PHA responses decreased in the terminal patients. The results do not support the idea that the methods currently available should be routinely used in the follow-up of cancer patients; rather, they indicate the need to seek new methods for this purpose.     

Impact of cutaneous IL-10 on resident epidermal Langerhans' cells and the development of polarized immune responses

Prolonged topical exposure of BALB/c mice to chemical contact and respiratory allergens stimulates, respectively, preferential Th1- and Th2-type responses with respect to serum Ab isotype and cytokine secretion phenotypes displayed by draining lymph node cells. We now report that differential cytokine secretion patterns are induced rapidly in the skin following first exposure to the contact allergen 2,4-dinitrochlorobenzene (DNCB) and the respiratory sensitizer trimellitic anhydride (TMA). TMA induced early expression of IL-10, a cytokine implicated in the negative regulation of Langerhans cell (LC) migration, whereas exposure to DNCB resulted in production of the proinflammatory cytokine IL-1beta. Associated with this, TMA provoked LC migration with delayed kinetics compared with DNCB, and local neutralization of IL-10 caused enhanced LC mobilization in response to TMA with concomitant up-regulation of cutaneous IL-1beta. We hypothesize that these differential epidermal cytokine profiles contribute to the polarization of immune responses to chemical allergens via effects on the phenotype of activated dendritic cells arriving in the draining lymph node. Thus, TMA-exposed dendritic cells that have been conditioned in vivo with IL-10 (a potent inhibitor of the type 1-polarizing cytokine IL-12) are effective APCs for the development of a Th2-type response.     

Nippostrongylus brasiliensis-induced depression of heterologous immune responses: effects of the histamine H2 receptor antagonist, cimetidine

Both mucosal and systemic immune responses are depressed in mice infected with the nematode Nippostrongylus brasiliensis and this is correlated with a striking increase in the numbers of histamine-containing, mucosal-associated mast cells. As suppressor T cells bear histamine H2 receptors, we have used the H2 antagonist, cimetidine, to test whether histamine mediates N. brasiliensis-induced immunodepression. In uninfected mice, mitogenic responses to PHA and Con A were increased by treatment with cimetidine; in some experiments this treatment also increased antibody responses to a T-dependent antigen (TNP-BGG). By contrast, these T- and B-cell responses were markedly depressed in mice infected with N. brasiliensis, and cimetidine treatment did not alter this parasite-induced immunodepression. These results imply that although histamine can modulate immune responses in uninfected animals, it is not a major component of the immunoregulatory pathway induced by infection with N. brasiliensis.     

Effects of immunotherapy and chemotherapy on immunocompetence. A study of patients with acute myeloblastic leukemia in remission

Summary The immunocompetence of 33 patients with acute myeloblastic leukemia in remission and treated with cytostatics (CT) was studied. In addition to cytostatics some of the patients were given immunotherapy (CT+IT).In an attempt to demonstrate immunization against allogeneic leukemic blast cells (or their extracts) or immunostimulation after immunotherapy or, alternatively, immunodepression after maintenance chemotherapy without immunotherapy, delayed hypersensitivity tests and lymphocyte stimulation tests were performed. In most cases PHA seemed to be a stronger stimulator than allogeneic lymphocytes and these seemed to be stronger than allogeneic blasts, although no difference was statisically significant.No significant differences were found in vitro or in vivo between the reactions of CT and CT+IT patients or their lymphocytes to allogeneic myeloblasts or to allogeneic lymphocytes. However, numerically, in vitro and in vivo CT+IT patients reacted more to myeloblasts, CT patients more to lymphocytes. This could suggest antigens on leukemic myeloblasts that are not found on lymphocytes. With present methods we could demonstrate neither immunodepression in patients given only chemotherapy nor nonspecific immunostimulation after immunotherapy. There was no significant difference between the two treatment groups in lymphocyte reactivity against PHA and allogeneic lymphocytes. Nor was the lymphocyte reactivity different from that in a group of healthy persons.Decreasing lymphocyte reactivity to PHA and allogeneic lymphocytes seemed to herald relapse.     

Decreased expression of autophagy-related proteins in malignant epithelial ovarian cancer

Some studies have shown that the change of autophagic capacity may correlate with malignant transformation. Our study was designed to investigate the expression and significance of two autophagy-related proteins, microtubule-associated protein 1 light chain 3 (LC3) and Beclin 1 in the tumorigenesis and development of epithelial ovarian carcinoma. We observed that the positive expression of LC3 and Beclin 1 was significantly higher in the samples of benign and borderline ovarian tumors than those in malignant epithelial ovarian cancers. The expression of LC3 and Beclin 1 was associated with FIGO stage and histological grade. No significant relationship was observed between age and histological grade. However, we observed that the expression of LC3 was not related to Beclin 1. Therefore, the decrease of autophagic capacity may be related to tumorigenesis and development of epithelial ovarian cancer.

Addendum: Shen Y, Liang LZ, Hong MH, Xiong Y, Wei M, Zhu XF. Expression and clinical significance of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in epithelial ovarian cancer. Ai Zheng 2008; 27:595-9.     

Decreased expression of autophagy-related proteins in malignant epithelial ovarian cancer

Some studies have shown that the change of autophagic capacity may correlate with malignant transformation. Our study was designed to investigate the expression and significance of two autophagy-related proteins, microtubule-associated protein 1 light chain 3 (LC3) and Beclin 1 in the tumorigenesis and development of epithelial ovarian carcinoma. We observed that the positive expression of LC3 and Beclin 1 was significantly higher in the samples of benign and borderline ovarian tumors than those in malignant epithelial ovarian cancers. The expression of LC3 and Beclin 1 was associated with FIGO stage and histological grade. No significant relationship was observed between age and histological grade. However, we observed that the expression of LC3 was not related to Beclin 1. Therefore, the decrease of autophagic capacity may be related to tumorigenesis and development of epithelial ovarian cancer.     

Mortality after cerebral angiography with or without radioactive Thorotrast: an international cohort of 3,143 two-year survivors.

There are few studies on the long-term sequelae of radionuclides ingested or injected into the human body. Patients exposed to radioactive Thorotrast in the 1930s through the early 1950s provide a singular opportunity, since the administration of this radiographic contrast agent resulted in continuous exposure to alpha particles throughout life at a low dose rate. We evaluated cause-specific mortality among an international cohort of 3,143 patients injected during cerebral angiography with either Thorotrast (n = 1,736) or a similar but nonradioactive agent (n = 1,407) and who survived 2 or more years. Standardized mortality ratios (SMRs) for Thorotrast and comparison patients were calculated, and relative risks (RR), adjusted for population, age and sex, were obtained by multivariate statistical modeling. Most patients were followed until death, with only 94 (5.4%) of the Thorotrast patients known to be alive at the closure of the study. All-cause mortality (n = 1,599 deaths) was significantly elevated among Thorotrast subjects [RR 1.7; 95% confidence interval (CI) 1.5-1.8]. Significantly increased relative risks were found for several categories, including cancer (RR 2.8), benign and unspecified tumors (RR 1.5), benign blood diseases (RR 7.1), and benign liver disorders (RR 6.5). Nonsignificant increases were seen for respiratory disease (RR 1.4) and other types of digestive disease (RR 1.6). The relative risk due to all causes increased steadily after angiography to reach a threefold RR at 40 or more years (P < 0.001). Excess cancer deaths were observed for each decade after Thorotrast injection, even after 50 years (SMR 8.6; P < 0.05). Increasing cumulative dose of radiation was directly associated with death due to all causes combined, cancer, respiratory disease, benign liver disease, and other types of digestive disease. Our study confirms the relationship between Thorotrast and increased mortality due to cancer, benign liver disease, and benign hematological disease, and suggests a possible relationship with respiratory disorders and other types of digestive disease. The cumulative excess risk of cancer death remained high up to 50 years after injection with >20 ml Thorotrast and approached 50%.     

肿瘤自身抗体检测对早期肺癌的诊断价值

目的探讨肺癌自身抗体P53、PGP9.5、SOX2、GAGE7、GBU4-5、MAGEA1和CAGE对早期肺癌的诊断价值。方法采用ELISA法检测肺癌组(46例首次确诊的肺癌患者)和对照组(包含22名肺部良性疾病患者及23例健康体检者)血清中肺癌自身抗体P53、PGP9.5、SOX2、GAGE7、GBU4-5、MAGEA1和CAGE表达水平,分析评价7种肺癌自身抗体单独检测和联合检测对肺癌的临床诊断价值。结果肺癌组患者血清中7种自身抗体的水平明显高于对照组(均P<0.05)。肺癌组患者7种抗体联合检测的阳性率明显高于对照组(65.22%vs 15.56%,χ²=23.252,P<0.001)。单一肺癌自身抗体检测的敏感性为4.35%~36.96%,特异性为93.30%~100.00%,AUC为0.695~0.832。7种肺癌抗体联合检测的敏感性为65.22%,特异性为84.44%,AUC为0.897。结论早期肺癌患者血清中7种肺癌自身抗体水平均较高。单一肺癌自身抗体检测的特异性较高,但敏感性较低。7种肺癌自身抗体联合检测具有较高的敏感性,可成为临床早期肺癌新的辅助诊断指标。     

Decon2LS: An open-source software package for automated processing and visualization of high resolution mass spectrometry data

Background

The majority of ovarian cancer biomarker discovery efforts focus on the identification of proteins that can improve the predictive power of presently available diagnostic tests. We here show that metabolomics, the study of metabolic changes in biological systems, can also provide characteristic small molecule fingerprints related to this disease.

Results

In this work, new approaches to automatic classification of metabolomic data produced from sera of ovarian cancer patients and benign controls are investigated. The performance of support vector machines (SVM) for the classification of liquid chromatography/time-of-flight mass spectrometry (LC/TOF MS) metabolomic data focusing on recognizing combinations or "panels" of potential metabolic diagnostic biomarkers was evaluated. Utilizing LC/TOF MS, sera from 37 ovarian cancer patients and 35 benign controls were studied. Optimum panels of spectral features observed in positive or/and negative ion mode electrospray (ESI) MS with the ability to distinguish between control and ovarian cancer samples were selected using state-of-the-art feature selection methods such as recursive feature elimination and L1-norm SVM.

Conclusion

Three evaluation processes (leave-one-out-cross-validation, 12-fold-cross-validation, 52-20-split-validation) were used to examine the SVM models based on the selected panels in terms of their ability for differentiating control vs. disease serum samples. The statistical significance for these feature selection results were comprehensively investigated. Classification of the serum sample test set was over 90% accurate indicating promise that the above approach may lead to the development of an accurate and reliable metabolomic-based approach for detecting ovarian cancer.     

Detection of aggressive prostate cancer associated glycoproteins in urine using glycoproteomics and mass spectrometry

Clinical management of prostate cancer remains a significant challenge due to the lack of available tests for guiding treatment decisions. The blood prostate‐specific antigen test has facilitated early detection and intervention of prostate cancer. However, blood prostate‐specific antigen levels are less effective in distinguishing aggressive from indolent prostate cancers and other benign prostatic diseases. Thus, the development of novel approaches specific for prostate cancer that can differentiate aggressive from indolent disease remains an urgent medical need. In the current study, we evaluated urine specimens from prostate cancer patients using LC‐MS/MS, with the aim of identifying effective urinary prostate cancer biomarkers. Glycoproteins from urine samples of prostate cancer patients with different Gleason scores were characterized via solid phase extraction of N‐linked glycosite‐containing peptides and LC‐MS/MS. A total of 2923 unique glycosite‐containing peptides were identified. Glycoproteomic comparison on urine and tissues from aggressive and non‐aggressive prostate cancers as well as sera from prostate cancer patients revealed that the majority of AG prostate cancer associated glycoproteins were more readily detected in patient's urine than serum samples. Our data collectively indicate that urine provides a potential source for biomarker testing in patients with AG prostate cancer.     

Uridine incorporation into the RNA of stimulated lymphocytes with different sets of sex chromosomes

Phytohemagglutinin (PHA)-induced stimulation of uridine incorporation into RNA was studied with cotton-wool isolated peripheral blood human lymphocytes. The amount of labeling in 24-hour cultures proved to be increased in men and in 45,XO patients, compared to women. With high doses of PHA applied, hyperreactivity of the 45, XO cells was revealed. PHA-activation in normal persons tends to decrease with advancing age. Complex genomic as well as sex hormone effects may be responsible for the differences observed.     

Rapid effects of BCG on T cell function in cancer patients

Summary Forty-four cancer patients were evaluated with multiple sequential tests to determine the effects of a single intracutaneous injection of BCG on the levels of total and high-affinity sheep erythrocyte-forming cells and on lymphoproliferative responses to a T cell mitogen (PHA) and antigens (MLC and PPD). The analysis of the data was complicated by considerable pretreatment variation in the results with some patients and by the lack of a consistent change in responses among all patients. However, exploratory statistical procedures made it possible to demonstrate that inoculation with BCG was associated with a significant transient increase, on day 3 post-injection, in levels of total and active rosette-forming cells and in lymphoproliferative responses to PPD. The effects on responses to PHA varied with the level of pretreatment reactivity with patients with initial low responses showing a significant increase on days 3 and 7 and patients with initial normal responses showing a significant decrease on days 3 and 7. Patients and normal individuals receiving no injection, and patients receiving isotonic saline injections, showed none of these changes. Thus it appears that BCG may cause measurable changes in T cell levels and lymphoproliferation to T cell mitogen (PHA) and antigen (PPD).     

Increased growth hormone responses to growth hormone releasing hormone and thyrotropin releasing hormone in patients with metastatic testicular cancer

In 16 patients with metastatic testicular cancer and 10 age matched male control subjects growth hormone (GH) responses to growth hormone releasing hormone (GHRH; 1 microgram/kg body weight iv.) and thyrotropin releasing hormone (TRH; 200 micrograms iv.) were measured. Basal GH levels and GH levels following stimulation with GHRH or TRH were significantly increased in cancer patients compared to control subjects. 9 patients with testicular cancer were studied both in the stage of metastatic disease and after they had reached a complete remission. In complete remission GH responses to GHRH tended to decrease but the differences did not reach statistical significance. Our data suggest an alteration of hypothalamic and/or pituitary regulation of GH secretion in patients with metastatic testicular cancer.     

Autophagy activity is associated with membranous sodium iodide symporter expression and clinical response to radioiodine therapy in non-medullary thyroid cancer

Although non-medullary thyroid cancer (NMTC) generally has a good prognosis, 30–40% of patients with distant metastases develop resistance to radioactive iodine (RAI) therapy due to tumor dedifferentiation. For these patients, treatment options are limited and prognosis is poor. In the present study, expression and activity of autophagy was assessed in large sets of normal, benign and malignant tissues and was correlated with pathology, SLC5A5/hNIS (solute carrier family 5 member 5) protein expression, and with clinical response to RAI ablation therapy in NMTC patients. Fluorescent immunostaining for the autophagy marker LC3 was performed on 100 benign and 80 malignant thyroid tissues. Semiquantitative scoring was generated for both diffuse LC3-I intensity and number of LC3-II-positive puncta and was correlated with SLC5A5 protein expression and clinical parameters. Degree of diffuse LC3-I intensity and number of LC3-II-positive puncta scoring were not discriminative for benign vs. malignant thyroid lesions. Interestingly, however, in NMTC patients significant associations were observed between diffuse LC3-I intensity and LC3-II-positive puncta scoring on the one hand and clinical response to RAI therapy on the other hand (odds ratio [OR] = 3.13, 95% confidence interval [CI] =1.91–5.12, P = 0.01; OR = 5.68, 95%CI = 3.02–10.05, P = 0.002, respectively). Mechanistically, the number of LC3-II-positive puncta correlated with membranous SLC5A5 expression (OR = 7.71, 95%CI = 4.15–11.75, P<0.001), number of RAI treatments required to reach remission (P = 0.014), cumulative RAI dose (P = 0.026) and with overall remission and recurrence rates (P = 0.031). In conclusion, autophagy activity strongly correlates with clinical response of NMTC patients to RAI therapy, potentially by its capacity to maintain tumor cell differentiation and to preserve functional iodide uptake.     

Cancer immunotherapy.

Important contributions that stimulated studies in cancer immunotherapy included: (1) the discovery of tumour-associated antigens; (2) the observation that infection with bacille Calmette-Guérin (BCG) in animals was protective against tumour challenge; and (3) the observation that immunodepression due either to malignant disease or to treatment of the disease, was, in some instances, related to prognosis. Immunotherapy trials with microbial agents have involved attempts to obtain a local effect by injecting the agent into the tumour or into the region of the tumour and to obtain a "systemic" effect distant from the site of injection. Trials with active specific immunotherapy involving tumour cells or tumour cell extracts have frequently involved the combination of these specific agents with a nonspecific adjuvant such as BCG. Recent studies with thymosin and levamisole in patients with lung cancer and other types of malignant disease have shown prolonged survival in the groups receiving immunotherapy.     

Non-Specific Factors that may influence Significance of Urinary Steroid Excretion in Breast Cancer

The urinary excretion of corticosteroids (17-oxogenic steroids) and adrenal androgens (11-deoxy-17-oxosteroids) was studied in women below the age of 50 in a variety of clinical situations for comparison with a normal group. The conditions studied were: chronic debility from non-malignant disease, weight reduction, admission to hospital and surgery for varicose veins, hepatic non-malignant disease, and non-mammary cancer.The objective of the study was to determine whether the changes found in early and advanced breast cancer and used to judge the prognosis of the disease are specific to the disease or are merely incidental to the degree of illness caused by the disease.Similar changes to those found in breast cancer—principally a reduction in the excretion of the androgens—were found in the women with severe hepatic disease and in advanced non-mammary cancer. These were also found to follow the effects of severe surgical stress.It is concluded that the changes found in breast cancer are a measure of the general systemic disturbance caused by the disease and are not due specifically to it. Nevertheless, the value of their prognostic significance remains unchallenged.     

Modification of immune competence by parasitic infections. I. Responses to mitogens and antigens in mice treated with Trichinella spiralis extract

Spleen cells from mice pretreated with a Trichinella spiralis extract (TsE-mice) showed severe depression of the response to lipopolysaccharide (LPS) and to concanavalin A (Con A), slight depression to phytohemagglutinin (PHA) and normal response to tuberculin purified protein derivative (PPD) as compared to saline-pretreated controls. Mice pretreated with bovine serum albumin (BSA-mice) revealed greatly reduced responses to LPS, somewhat reduced response to Con A, and normal responses to PHA and to PPD. Only TsE-mice showed significant reduction in the number of rosette-forming cells and of direct and indirect plaque-forming cells (DPFC and IPFC). BSA-mice exhibited some reduction of the DPFC only. Direct hemagglutinating (HA) titers were equivalent in the 3 groups after immunization with sheep erythrocytes but facilitated HA titers were depressed in TsE-mice. The total number and the number of viable cells were similar in the spleens of all animals. TsE treatment causes a reduction in the number of T1 lymphocytes and an inhibition of the late differentiation of B cells in the spleen. Suppressor T-cells apparently play a major but not exclusive role in T. spiralis-induced nonspecific immunodepression.     

Diminished lymphocyte responses to phytohemagglutinin in lung cancer

The PHA response of blood lymphocytes from lung cancer patients was found to be diminished in comparison to normal. Sera from these patients inhibited the blastogenic response of blood lymphocytes from normal subjects. Normal sera could restore to various levels the diminished PHA response of lymphocytes from lung cancer patients. The results suggest that the immunosuppression seen in lung cancer may be mediated by a factor (s) in the serum which might bound reversibly to a certain subpopulation of T-cells and permanently to another and or some other inhibitory mechanism does exist.