COMBINED EFFECT OF ANTIBODIES TO BENZO[A]PYRENE, ESTRADIOL AND PROGESTERONE UPON SEX HORMONE CONCENTRATIONS IN BLOOD SERUM OF PREGNANT WOMEN WITH CONGENITAL MALFORMATIONS OF FETUS

Specific antibodies against environmental chemical gene toxicants and endogenous steroid hormones are shown to modulate concentrations of these compounds in blood serum and their biological effects in experimental models. However, probable hazards of such antibodies in human teratogenesis are still unknown. In particular, potential correlations between specific serum antibodies, sex hormone levels in pregnant women, and congenital malformations in newborns are not clear. The aim of this study was to identify possible associations between occurrence of antibodies to benzo[a]pyrene, estradiol and progesterone (Bp, Es and Pg, respectively), and congenital malformations, and effects of these antibodies upon Es and Pg concentrations in blood serum of pregnant women. We have included into the study 182 women with normal pregnancy and 101 females with congenital malformations of fetus. A non-competitive solid phase immunoassay was performed using Bp, Es and Pg conjugated to bovine serum albumin as antigens. Es and Pg serum concentrations were measured using immunoassay test-systems of “Immunotech” (Moscow). Results: strong positive correlations were revealed between the levels of studied antibodies in the both groups. High IgA-Bp/IgA-Es (> 3) and IgA-Bp/IgA-Pg ratios (> 3) were associated with congenital malformations (OR = 2.2, p = 0.013 and OR = 6.8, p < 0.0001). Positive correlations were revealed between Pg/Es and IgA-Bp/IgA-Es (rS = 0.62, p < 0.0001), and IgA-Bp/IgA-Pg ratios (rS = 0.77, p < 0.0001) in cases with inborn malformations. Similar correlations were found for the women who had normal pregnancy (rS = 0.4, p = 0.0001, and rS = 0.23, p = 0.026, respectively). The Pg/Es proportion correlated with IgG-Bp/IgG-Es (rS = 0.46, p = 0.002), and with IgG-Bp/IgG-Pg ratio (rS = 0.5, p = 0.0009) in cases of malformations, but not in women with normal pregnancy. Conclusion: we have revealed novel associations between congenital malformations of fetus and ratios of IgA-Bp/IgA-Es, as well as IgA-Bp/IgA-Pg, like as positive correlations between hormonal Pg/Es proportions, and ratios of specific antibodies in pregnant women

IMMUNOLOGICAL IMBALANCE IN BREAST CANCER AND LUNG CANCER IN POSTMENOPAUSAL WOMEN

Previous studies reported some associations between class A antibodies specific for benzo[a]pyrene (IgA-Bp), estradiol (IgA-Es) and progesterone (IgA-Pg) and breast cancer (BC) in women like as with lung cancer (LC) in men. It was suggested that IgA-Bp and IgA-Es may stimulate tumor initiation and promotion, whereas IgA-Pg may inhibit the in vivo human carcinogenesis.The purpose of this study was to identify the suggested associations of such immunological imbalance with BC and LC in postmenopausal women.The serum A-class antibodies specific to benzo[a]pyrene, estradiol and progesterone (IgA-Bp, IgA-Es, IgA- Pg) were studied in 335 healthy women, 824 breast cancer (BC) patients and 127 cases of lung cancer (LC) by means of non-competitive solid phase immunoassay. The following results were obtained: Increased ratio of IgA-Bp and IgA-Es amounts exceeding the IgA-Pg levels was associated with a higher risk of breast cancer (OR = 2.8 and 2.4 respectively, p < 0.0001), and higher risk of LC (OR = 2.9 and 2.8, respectively, p < 0.0001). Conversely, the OR values decreased to 0.3-0.4 for BC and LC if IgA-Pg levels were higher than IgA-Bp and IgA-Es levels (p < 0.0001). These findings confirm the hypothesis that IgA-Bp and IgA-Es are capable to stimulate, and IgA-Pg, to inhibit the BC and LC occurrence n postmenopausal women. The balance between IgA-Bp and IgA-Es, on the one hand, and IgA-Pg, on the other hand, is much more important than individual contents of these antibodies.In conclusion, the phenomenon of “immunological interference” is revealed, i.e., the mutual enhancement of IgA-Bp and IgA-Es effects, thus, probably, stimulating the initial and subsequent events of carcinogenesis initiation and promotion, with a weak anticancer effect of IgA-Pg, and by weakening the mutual procarcinogenic effects of IgA-Bp and IgA-Es by the marked effect of IgA-Pg

IMMUNOREGULATION OF BLOOD SERUM ESTRADIOL AND PROGESTERONE LEVELS IN POSTMENOPAUSAL WOMEN

Specific antibodies against estradiol (Es) and progesterone (Pg) are known to modulate blood serum concentrations of these hormones and their biological effects after immunization of animals. It was suggested that specific IgA-Es and IgA-Pg could influence on Es and Pg levels in human blood serum. The purpose of this study was to identify the suggested correlations between serum Es and Pg and specific IgA-Es and IgA-Pg in postmenopausal healthy women (HW) and breast cancer patients (BCP). The serum levels of Es, Pg, IgA-Es and IgA-Pg were studied in 226 HW and 633 BCP by means of solid-phase immunoassay. The following results were obtained. The levels of Es in BCP (0.25 nmol/l) were higher than in HW (0.16; р < 0.0001). The levels of Pg were lower (0.79 vs 0.87; р < 0.0001), and individual Pg/Es ratios were lower (3.19 vs 6.64; р < 0.0001). Individual IgA-Pg/IgA-Es ratios correlated with decrease of Es (rs = -0.15; p = 0.029), with increase in Pg (rs = 0.38; р < 0.0001), and with increased Pg/Es ratio (rs = 0.29; р < 0.0001) in healthy women. Similar correlations were determined in BCP (correspondingly: rs = -0.14, р < 0.001; rs = 0.1, р = 0.009; rs = 0.15, р < 0.0001). The decrease of Es and increase of Pg and Pg/Es in BCP were less significant than in HW: the a quotients in regression у = ах+b (y = hormones levels and x = antibodies levels) in BCP were 3 to 4-fold lower than in HW. These peciliarities of interrelations between hormones and specific antibody levels were revealed only in ER+/PR+ BCP but not in ER+/PR- and ER-/PR- BCP. In conclusion, we have confirmed a suggestion about participation of specific antibodies in regulation of steroids levels in human blood serum. The immune regulation of hormonal status was weakened in BCP

Immunological imbalance, gene polymorphism of biotransformation enzymes, and steroid hormone receptors in tumors in breast cancer patients

It is well known that results of breast cancer (BC) hormonal therapy depend on expression of tumor estradiol and progesterone receptors (ER and PR) in tumor tissue. Mechanisms of ER+/PR+ tumors conversion to ER+/PR- and ER-/PR- tumors remain scarcely studied. The decrease of steroid receptors expression seems to depend on action of genotoxic metabolites of environmental carcinogens (particularly, benzo[a]pyrene, BP) and endogenous steroids (in particular, estradiol, E2). The formation of these metabolites is regulated by the biotransformation enzymes. On the other hand, the formation of DNA-adducts with genotoxic metabolites may induce the synthesis of specific antibodies. Previously, it was shown that increase of the serum IgA-antibodies levels against Bp and E2 over the levels of IgA-antibodies against progesterone (IgA-Bp/IgA-Pg > 1 and IgA-E2/IgA-Pg), could be interpreted as immunological imbalance associated with high BC risk in healthy women. The purpose of this study was to detect the suggested associations between ER+/PR+ tumors conversion to ER+/PR+ and ER-/PR- tumors and immunological imbalance in the BC patients with distinct gene variants of biotransformation enzymes: CYP1A1*2A (rs 4646903), CYP1B1 (rs1056836), CYP19A1 (rs2470152), GSTT1 (del), GSTP1 (rs1695). The IgA-Bp, IgA-E2 and IgA-Pg were studied in 1321 non-smoking BC patients by non-competitive solid phase immunoassay. The conjugates of Bp, E2 and Pg with bovine serum albumin were adsorbed as target antibodies. The goat antibodies against human IgA conjugated with horseradish peroxidase were used for detection of the studied specific antibodies. Gene polymorphisms of biotransformation enzymes were analyzed by the real-time PCR. Tumor ER and PR were detected by the standard immunohistochemical methods.ER+/PR+ tumors in BC patients at the stage I (N = 534) were found in 68.7%, ER+/PR- in 15.6%, ER-/ PR- in 15.7%. In BC patients at the II-IV stage (N = 787), frequency of ER+/PR+ tumors decreased to 60.2%, ER+/PR- was similar (15.8%), and ER-/PR- increased to 24.0% (p < 0.0001). These alterations were revealed in BC patients at the IgA-Bp/IgA-Pg ratios > 1, and IgA-E2/IgA-Pg > 1 only. There were no differences found between BC patients at stage I and II-IV at the ER+/PR+, ER+/PR-, ER-/PR- frequencies when these ratios were low.The frequency of ER+/PR+ tumors in homozygotes TT of CYP19A1 was 77.1% at the I stage and 60.1% at the II-IV stages. Respectively the frequencies of ER-/PR- tumors were 11.8% and 26.1% (p < 0.001). ER+/ PR+ tumors were revealed in GSTT1 “+” BC patients at the I stage in 68.7% and at the II-IV stages in 58.0%. Respectively ER-/PR- tumors were found in 16.6% and 24.5% (p < 0.0004). The frequency of ER+/PR+ tumors was 57.1% in homozygotes GG of GSTP1 at the I stage and 60.7% at the II-IV stages. Respectively the frequencies of ER+/PR- were 14.3% and 22.2% and ER-/PR- were 28.6% and 19.0% (p < 0.001). Proportions of low and high IgA-Bp/IgA-Pg and IgA-E2/IgA-Pg ratios were the same at the any enzyme genotype of studied CYP or GST variants. In conclusion, we have revealed a sufficient contribution of immunological imbalance to the conversion of steroid receptors in breast cancer growth, being independent of several CYP and GST gene polymorphisms

Association of antibodies to benzo[a]pyrene, estradiol and progesterone with gene polymorphisms of cytokines in postmenopausal women

Previous studies reported some associations between IgA and IgG antibodies specific to benzo[a] pyrene (Bp), estradiol (Es) and progesterone (Pg), and breast cancer (BC) in postmenopausal women. Likewise, the individual ratios of these antibodies (IgA-Bp/IgA-Pg, IgG-Bp/IgG-Pg, IgG-Es/IgG-Pg, IgG-Es/IgG-Pg) were associated with BC. It was suggested that development of antibodies to chemical carcinogens and steroid hormones was determined by functional polymorphisms of cytokine genes. The purpose of this study was to identify the suggested associations of antibodies to Bp, Es, Pg, and their individual ratios to the following gene polymorphisms: IL1RN (rs4251961), IL1B (rs16944), IL6 (rs1800795, rs1800796, rs1554606), IL8 (rs4073), TNFA (rs1800629) and CD40 (rs6074022) detected in postmenopausal healthy women and BC patients.The serum IgA and IgG antibodies specific to Bp, Es and Pg were studied in 470 healthy women and 995 BC patients by non-competitive solid phase immunoassay. The conjugates of Bp, Es, Pg with bovine serum albumin were used as adsorbed antigen. The goat antibodies against human IgA or IgG conjugated with horseradish peroxidase were used for the detection of bound hapten-specific antibodies. Cytokine gene polymorphisms were analyzed by the real-time PCR.Associations between the studied antibodies and their ratios with the gene polymorphisms in IL1RN (rs4251961), IL6 (rs1800795), TNFA (rs1800629) and CD40 (rs6074022) were found in healthy women. Higher individual ratios of IgA-Bp/IgA-Pg (p = 0.0001), IgG-Bp/IgG-Pg (p < 0.0001), IgG-Es/IgG-Pg (p = 0.0003) were associated with the allele C gene IL1RN. The higher IgG-Es levels were more common in the persons with allele G gene IL6 (p = 0.007), and with C allele of CD40 gene (p = 0.005). The high IgA-Pg levels were associated with A allele gene of TNFA (p = 0.008). Associations of antibodies were found only with genes polymorphisms in CD40 (rs6074022) in BC patients. Higher IgG-Es levels were more common in persons with allele T gene CD40 (p = 0.007).In conclusion, we revealed the participation of cytokines in immune regulation of antibody genesis for environmental chemical carcinogens and endogenous steroid hormones in healthy women and BC patients. The future investigations of antibodies specific to Bp, Es and Pg combined with the analysis genes polymorphisms in cytokines will be useful for detection of the individual hormone-dependent cancer risks in humans

THE ASSOCIATION OF IMMUNE RESPONSE TO XENOAND ENDOBIOTIKS AND THEIRS BIOTRANSFORMATION ENZYME GENE POLYMORPHISM WITH CONGENITAL MALFORMATIONS OF THE FETUS

The correlation between immune response to benzo(a)pyrene and progesterone in combination with detoxification enzyme gene polymorphism (CYPIA2*1F and GSTTI) and the occurrence of congenital malformations of the fetus was revealed. It is shown that at high ratio of IgA-antibodies to benzo(a)pyrene and to progesterone in conjunction with the mutant allele of the gene CYPIA2*1F and the deletion genotype of the gene GSTTI chance of the reproductive pathology developing increases to 41 times

Антитела класса G, специфичные к бензо[а]пирену, эстрадиолу и прогестерону у женщин с колоректальным раком и раком молочной железы

The study was aimed to determine the level of class G antibodies specific to Bp, Es, Pg (IgG-Bp, IgG-Es, IgG-Pg) in patients with colorectal and breast cancers. Material and methods. The content of these antibodies in the blood serum of healthy women (n=401), patients with colorectal cancer (n=219) and patients with breast cancer (n=1469) was studied using a non-competitive enzyme immunoassay. Statistical analysis of the results was performed using the Statistica 8.0 software. Results. The levels of IgG-Bp >7 and IgG-Es>6 were higher in patients with colorectal cancer than in healthy controls (66 % vs 25 %, p<0.0001, OR=5.9 and 58 % vs s 45 %, p=0.002, OR=1.7, respectively). The individual antibody ratios of IgG-Bp/IgG-Es >1, IgG-Bp/IgG-Pg>1.5, and IgG-Es/IgG-Pg>1.5 were also higher in patients with colorectal cancer than in healthy controls (74 % vs 34 %, p<0.0001, OR=5.6; 75 % vs 28 %, p<0.0001, and 58 % vs 38 %, p<0.0001, OR=2.3, respectively). Compared to healthy controls, breast cancer patients had higher values of IgG-Bp >6 (57 % vs 33 %, p<0.0001, OR=2.7) and IgG-Es>5 (62 % vs 53 %, p=0.003, or=1.4) and ratios of IgG-Bp/IgG-Es>1 (55 % vs 34 %, p<0.0001, or=2.4), IgG-Bp/IgG-Pg>1.3 (71 % vs 36 %, p<0.0001, or=4.5) and IgG-Es/IgG-Pg>1.4 (62 % vs 44 %, p<0.0001, or=2.1). Compared to breast cancer patients, colorectal cancer patients had higher values of IgG-Bp>7 (66 % vs 50 %, p<0.0001) and the ratios of igg-Bp/IgG-Es >1 (74 % vs 55 %, p<0.0001) and IgG-Bp/IgG-Pg>1.5 (76 % vs 60 %, p<0.0001). Conclusion. IgG-Bp, IgG-Es, and IgG-Pg immunoassay could serve as a screening tool to identify population at risk of colorectal and breast cancers.Цель исследования ‒ определение уровня антител класса G, специфичных к бензо[а]пирену (IgG-Bp), эстрадиолу (IgG-Es) и прогестерону (IgG-Pg) у больных колоректальным раком и раком молочной железы. Материал и методы. С помощью неконкурентного иммуноферментного анализа исследовали содержание этих антител в сыворотке крови здоровых женщин (n=401), больных колоректальным раком (n=219) и раком молочной железы (n=1469). Статистический анализ результатов проводили с помощью программы Statistica 8.0. Результаты. У больных колоректальным раком по сравнению со здоровыми лицами чаще встречались высокие значения IgG-Bp >7 (66 % vs 25 %, p<0,0001, OR=5,9) и IgG-Es >6 (58 % vs 45 %, p=0,002, OR=1,7), а также индивидуальные соотношения антител: IgG-Bp / IgG-Es >1 (74 % vs в 34 %, p<0,0001, OR=5,6); IgG-Bp / IgG-Pg >1,5 (75 % vs 28 %, p<0,0001, OR=7,8); IgG-Es / IgG-Pg >1,5 (58 % vs 38 %, p<0,0001, OR=2,3). У больных раком молочной железы по сравнению со здоровыми лицами чаще встречались высокие значения IgG-Bp >6 (57 % vs 33 %, p<0,0001, oR=2,7) IgG-Es >5 (62 % vs 53 %, p=0,003, OR=1,4), а также соотношения IgG-Bp/ IgG-Es >1 (55 % vs 34 %, p<0,0001, OR=2,4), IgG-Bp / IgG-Pg >1,3 (71 % vs 36 %, p<0,0001, OR=4,5) и IgG-Es / IgG-Pg >1,4 (62 % vs 44 %, p<0,0001, OR=2,1). У больных колоректальным раком по сравнению с больными раком молочной железы чаще встречались высокие значения IgG-Bp >7 (66 % vs 50 %, p<0,0001), а также соотношения IgG-Bp / IgG-Es >1 (74 % vs 55 %, p<0,0001) и IgG-Bp IgG-Pg >1,5 (76 % vs 60 %, p<0,0001). Заключение. Иммуноанализ IgG-Bp, IgG-Es и IgG-Pg можно использовать для создания диагностических тест-систем колоректального рака и рака молочной железы

ANTIBODIES AND ANTI-ANTIBODIES TO STEROID HORMONES, AND BREAST CANCER RISK

Antibodies against sex steroid hormones are known to modulate their serum concentration and  inhibit experimental breast cancer (BC). Hence, these effects could be changed by according anti-idiotypic  antibodies. However, relationships between antibodies and anti-idiotypic antibodies and BC in women are still  poorly studied. The aim of this study was to identify possible associations between occurrence of antibodies  against estradiol and progesterone (IgG-Es1and IgG-Pg1) and according antiidiotypic antibodies (IgG-Es2  and IgG-Pg2), and postmenopausal BC.  Eighty-nine  healthy  women  and  273  BC  patients  were  examined.  A  non-competitive  solid  phase  immunoassay for IgG-Es1 and IgG-Pg1 was performed using estradiol and progesterone conjugates with bovin  serum albumin as antigens. Monoclonal antibodies against Es and Pg as antigens have been used for noncompetitive solid phase immunoassay of IgG-Es2 and IgG-Pg2.  Results: absence of both IgG-Es1 and IgG-Pg1 was revealed in 53.9% of healthy donors and 41.0% of  BC patients (p = 0.04; OR = 0.6). Presence of IgG-Es1 without IgG-Pg1, or IgG-Pg1 without IgG-Es1was  detected for, respectively, 15.7% and 20.2% of healthy women, and in 10.3% and 7.7% of BC patients (p > 0.05).  Simultaneous increase of both IgG-Es1 and IgG-Pg1 was revealed in 10.1% of healthy donors and in 41.0%  of BC patients (p < 0.0001; OR = 5.3). Absence of a single antibogy (IgG-Es2 or IgG-Pg2) showed similar  frequency in the groups under study. Simultaneous increase of both IgG-Es2 and IgG-Pg2 was detected in  52.8% of healthy women and 34.8% of BC patients (p = 0.03; OR = 0.4). Conclusion: We have first revealed  an immunostimulating synergistic effect of antibodies against sex steroid hormones and immunoinhibitory  synergistic effect of appropriate anti-idiotypic antibodies upon the postmenopausal breast cancer risk