The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival

Background: The serum levels of IgG antibodies reactive to glycoconjugates (TF, Tn and αGal) were found to be associated with prognosis of gastrointestinal cancer patients. Aim: To study the relation between the levels of serum antibodies to TF-pAA, Tn-PAA and αGal-PAA polyacrylamide-based glycoconj...

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Published in:Experimental Oncology
Date:2016
Main Authors: Smorodin, E.P., Sergeyev, B.L.
Format: Article
Language:English
Published: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2016
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Cite this:The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival / E.P. Smorodin, B.L. Sergeyev // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 117-121. — Бібліогр.: 31 назв. — англ.

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spelling Smorodin, E.P.
Sergeyev, B.L.
2018-06-17T20:26:46Z
2018-06-17T20:26:46Z
2016
The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival / E.P. Smorodin, B.L. Sergeyev // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 117-121. — Бібліогр.: 31 назв. — англ.
1812-9269
https://nasplib.isofts.kiev.ua/handle/123456789/137989
Background: The serum levels of IgG antibodies reactive to glycoconjugates (TF, Tn and αGal) were found to be associated with prognosis of gastrointestinal cancer patients. Aim: To study the relation between the levels of serum antibodies to TF-pAA, Tn-PAA and αGal-PAA polyacrylamide-based glycoconjugates and survival in breast cancer. Materials and Methods: The preoperative level of IgG antibodies was analysed in the serum of patients (n = 59) using ELISA with polyacrylamide-glycoconjugates namely, TF-pAA (amide-type), and ethanolamide-conjugates Tn-PAA and αGal-PAA. Survival rate and hazard ratio (HR) were assessed by the Kaplan — Meier method and Cox univariate analysis in different pathomorphological groups. Results: Significantly better survival was observed in patients with an increased level of anti-TF-pAA antibodies both for all patients in total and groups in stages II–III; N1–2 and G3 (p = 0.008–0.021, HR = 0.18–0.23, mean survival time in months 164–186 vs 69–121). A trend to worse survival was observed in increased level of anti-Tn IgG (stages II–III) and anti-αGal IgG (G3): p = 0.075, HR = 2.49 and p = 0.066, HR = 3.27, respectively. Conclusion: The method for the determination of circulating anti-TF-pAA IgG may be a useful supplement in long-term prognostic assessment of patients with breast cancer.
This study was supported by a grant № 8399 from the Estonian Science Foundation.
en
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
Experimental Oncology
Original contributions
The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
Article
published earlier
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
title The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
spellingShingle The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
Smorodin, E.P.
Sergeyev, B.L.
Original contributions
title_short The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
title_full The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
title_fullStr The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
title_full_unstemmed The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
title_sort level of igg antibodies reactive to tf, tn and alpha-gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival
author Smorodin, E.P.
Sergeyev, B.L.
author_facet Smorodin, E.P.
Sergeyev, B.L.
topic Original contributions
topic_facet Original contributions
publishDate 2016
language English
container_title Experimental Oncology
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
format Article
description Background: The serum levels of IgG antibodies reactive to glycoconjugates (TF, Tn and αGal) were found to be associated with prognosis of gastrointestinal cancer patients. Aim: To study the relation between the levels of serum antibodies to TF-pAA, Tn-PAA and αGal-PAA polyacrylamide-based glycoconjugates and survival in breast cancer. Materials and Methods: The preoperative level of IgG antibodies was analysed in the serum of patients (n = 59) using ELISA with polyacrylamide-glycoconjugates namely, TF-pAA (amide-type), and ethanolamide-conjugates Tn-PAA and αGal-PAA. Survival rate and hazard ratio (HR) were assessed by the Kaplan — Meier method and Cox univariate analysis in different pathomorphological groups. Results: Significantly better survival was observed in patients with an increased level of anti-TF-pAA antibodies both for all patients in total and groups in stages II–III; N1–2 and G3 (p = 0.008–0.021, HR = 0.18–0.23, mean survival time in months 164–186 vs 69–121). A trend to worse survival was observed in increased level of anti-Tn IgG (stages II–III) and anti-αGal IgG (G3): p = 0.075, HR = 2.49 and p = 0.066, HR = 3.27, respectively. Conclusion: The method for the determination of circulating anti-TF-pAA IgG may be a useful supplement in long-term prognostic assessment of patients with breast cancer.
issn 1812-9269
url https://nasplib.isofts.kiev.ua/handle/123456789/137989
citation_txt The level of IgG antibodies reactive to TF, Tn and Alpha-Gal polyacrylamide-glycoconjugates in breast cancer patients: relation to survival / E.P. Smorodin, B.L. Sergeyev // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 117-121. — Бібліогр.: 31 назв. — англ.
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AT smorodinep levelofiggantibodiesreactivetotftnandalphagalpolyacrylamideglycoconjugatesinbreastcancerpatientsrelationtosurvival
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first_indexed 2025-11-25T12:45:32Z
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fulltext Experimental Oncology ��� �������� ���� ���ne���� �������� ���� ���ne� ���ne� ��� THE LEVEL OF IgG ANTIBODIES REACTIVE TO TF, Tn AND ALPHA- Gal POLYACRYLAMIDE-GLYCOCONJUGATES IN BREAST CANCER PATIENTS: RELATION TO SURVIVAL E.P. Smorodin*, B.L. Sergeyev Department of Oncology and Immunology, National Institute for Health Development, Hiiu 42, Tallinn 11619, Estonia Background: The serum levels of IgG antibodies reactive to glycoconjugates (TF, Tn and αGal) were found to be associated with prognosis of gastrointestinal cancer patients. Aim: To study the relation between the levels of serum antibodies to TF-pAA, Tn-PAA and αGal-PAA polyacrylamide-based glycoconjugates and survival in breast cancer. Materials and Methods: The preoperative level of IgG antibodies was analysed in the serum of patients (n = 59) using ELISA with polyacrylamide-glycoconjugates namely, TF-pAA (amide-type), and ethanolamide-conjugates Tn-PAA and αGal-PAA. Survival rate and hazard ratio (HR) were assessed by the Kaplan — Meier method and Cox univariate analysis in different pathomorphological groups. Results: Significantly better survival was observed in patients with an increased level of anti-TF-pAA antibodies both for all patients in total and groups in stages II–III; N1–2 and G3 (p = 0.008–0.021, HR = 0.18–0.23, mean survival time in months 164–186 vs 69–121). A trend to worse survival was observed in increased level of anti-Tn IgG (stages II–III) and anti-αGal IgG (G3): p = 0.075, HR = 2.49 and p = 0.066, HR = 3.27, respectively. Conclusion: The method for the determination of circulating anti-TF-pAA IgG may be a use- ful supplement in long-term prognostic assessment of patients with breast cancer. Key Words: breast cancer, survival, glycoconjugates, antibodies, microbiota, dysbiosis. INTRODUCTION Resident microbiota in h�mans colonizes bo�ndary s�rfaces maintaining imm�ne homeostasis [�]. Imba­ lance in microbial comm�nities� or microbial dysbiosis� has been shown to contrib�te to the cancero�s patho­ genesis and progression incl�ding breast cancer [�� �]. Microbial glycans are key antigens in host­microbial interactions [4]. Blood gro�p related cryptic glycans Thomsen — Friedenreich �TF� Galβ���GalNAcα�� Tn �GalNAcα� and αGal �Galα���Galβ� and/or mimicking determinants are present in enteric bacteria [5��]. Besides� TF and Tn are t�mor­associated glycans �TAG� beca�se they are freq�ently expressed in adenocarci­ nomas. The increased expression of TF and Tn is asso­ ciated with the t�mor invasion and metastases [����]. The αGal epitope is aberrantly expressed in h�man cells and it may be involved in a�toimm�ne diseases [�]. Be­ ing a constant so�rce of foreign antigens� microbiota stim�lates the prod�ction of anti­glycan antibodies. It is considered that the nat�ral anti­TF� ­Tn and ­αGal antibodies in h�man are ind�ced by enteric microorga­ nisms [5� �]. The high IgG antibody level and its change may reflect the adaptive imm�ne response of the host to microorganisms or increased a�toreactivity to self an­ tigens. Comparative data abo�t spontaneo�sly occ�r­ ring anti­glycan antibodies in cancer� their a�toreactivity to “self” and “altered self” antigens incl�ding TAG de­ serve a circ�mstantial investigation. We demonstrated the prognostic significance of the level of ser�m IgG antibodies to some glycans� and its relation to clinical parameters� t�mor progression and histopathological grading in gastrointestinal cancer patients [�����]. The antibody response to microbial glycans and its rela­ tion to s�rvival has not been chara cterized in patients with breast cancer yet. The aim of the present st�dy was to eval�ate whe­ ther the level of IgG antibodies to TF­pAA� Tn­PAA and αGal­PAA is related to s�rvival of patients with breast cancer� taking also into consideration the stage of the disease� t�mor vol�me� morphology and the t�mor spread in regional lymph nodes. MATERIALS AND METHODS Patients. The investigation was carried o�t in ac­ cordance with the ICH GCP Standards and approved by Tallinn Medical Research Ethics Committee� Esto­ nia. The informed consent was obtained from each patient �nder examination. Females with breast cancer in stages I�III �n = 59� whose diagnosis was verified by the pTNM system [�4]� were incl�ded in the st�dy. The chemo­ and X­ray therapy­prescribed patients as well as patients who died of ca�ses other than t�mor rec�rrence were excl�ded from the st�dy. The median and mean age ± SD was 5�.5 and 5�.� ± 9.4. Glycoconjugates. The synthetic glycoconj�gates that are homogeno�s antigens with a single reiterative glycotope [�5] were obtained from Lectinity Holding Inc. The sol�ble TF conj�gate with N­�ns�bstit�ted polyacrylamide �TF­pAA� amide­type� as well as etha­ nolamide­type glycoconj�gates of the poly[N­��­ hydroxyethyl�acrylamide] namely� Tn­PAA and αGal­ PAA �Bdi­PAA� were �sed in ELISA. The TF­pAA con­ tained �.� mol of TF per � mol of pAA. The rest of the conj�gates had �.� mol of a saccharide per � mol Submitted: April 05, 2016. *Correspondence: Tel.: +372-6593934; fax: +372-6593901; E-mail: jevgeni.smorodin@tai.ee Abbreviations used: HR — hazard ratio; IEF — isoelectric focusing; PAA — polyacrylamide; R — responders; WR — weak responders; TAG — tumor-associated glycans; TF — Thomsen — Friedenreich glycan; Tn — GalNAcα; αGal — (Galα1–3Galβ). Exp Oncol ���� ��� �� ������� ��� Experimental Oncology ��� �������� ���� ���ne� of PAA. Tris�hydroxymethyl�aminomethane­PAA was �sed as an adeq�ate negative control [��]. Indirect ELISA. The assay was performed as de­ scribed earlier [��]. The sera were kept at 4 °C for no longer than three weeks or were frozen �−5� °C� and thawed once before �se. The IgG antibody level was estimated as a ratio of Atest/Acontrol� where Atest is the absorbance with the glycoconj�gate and Acontrol� with the Tris­PAA. The variation coefficient was �%. Isoelectric focusing (IEF) and Western blotting. IEF was performed in a �% agarose IEF �GE Healthcare Bio­Sciences AB� Sweden� containing ��% glycerol as described in [��]. A pH gradient was formed in the gel with 4% Ampholytes high resol�tion ���� �Sigma­ Aldrich� USA� and �% Pharmalyte 5�� �GE Healthcare Bio­Sciences AB� Sweden�. Statistical analysis. The levels of antibodies were st�died in relation to overall s�rvival time starting from the date of the preoperatively taken blood sample. The Kaplan — Meier method was �sed for the esti­ mation of s�rvival c�rves in the �nivariate analysis of patients gro�ps� the significance of differences was analyzed by the log­rank test. The Cox proportional hazards model was �sed to eval�ate the risk of death. Statistical analysis was performed �sing SPSS� ver­ sion ��.�. The median �M� of the level of anti­TF­pAA� ­Tn and ­αGal antibodies was �sed as c�t­off to dis­ criminate between responders �R� the level above or eq�al to M� and weak responders �WR� the level below M�. S�rvival of R vs WR was eval�ated in s�b­ gro�ps divided by stages� t�mor vol�me� t�mor grade and regional lymph node metastases. The difference in s�rvival between patient gro�ps was considered to be significant when p ≤ �.�5. RESULTS Median and mean of the preoperative antibody le vels shown in Table � ranked as TF­pAA<Tn<αGal. Similar ratio of antibody levels was observed in gastro­ intestinal cancer patients and donors. The TF­R of dif­ ferent gro�ps demonstrated a significantly better s�r­ vival: p < �.�5� hazard ratio �HR� �.����.��� mean s�rvival time of R > WR �Table �; Fig. �� a�c�. The dif­ ference in s�rvival between TF­R and TF­WR in stages I�II and N� as well as in grades G��� or t�mor vo­ l�me T��4 was insignificant. The difference in s�r­ vival between Tn­R and Tn­WR as well as αGal­R and αGal­WR was insignificant for all patients or their patho­ morphological s�bgro�ps. A trend to a worse s�rvival of Tn­R and αGal­R �HR �.4���.��� in some s�bgro�ps was observed �Table �; Fig. �� d�. Table 1. Median and mean of the antibody level in the serum of patients Antibodies Median Mean ± SD, R Mean ± SD, WR TF-pAA 1.255 2.999 ± 2.463 1.134 ± 0.063 Tn 1.805 3.913 ± 2.487 1.380 ± 0.236 αGal 3.790 6.293 ± 2.858 2.180 ± 0.790 The specificity of anti­TF­pAA antibodies affinity­ isolated from cancero�s sera was described ear­ lier [��]. In the present st�dy� we analyzed antibodies �sing IEF and the IgG­Western blotting and showed their polyclonal pattern �Fig. ��. 0 20 40 60 80 100 0 50 100 150 200 Su rv iva l p ro ba bi lit y (% ) Months HR = 0.23 P = 0.021 0 20 40 60 80 100 0 50 100 150 200 Su rv iva l p ro ba bi lit y (% ) Months HR = 3.27 P = 0.066 0 20 40 60 80 100 0 50 100 150 200 Su rv iva l p ro ba bi lit y (% ) Months HR = 0.18 P = 0.017 0 20 40 60 80 100 0 50 100 150 200 Su rv iva l p ro ba bi lit y, % Months HR = 0.22 P = 0.012 d b c a Fig. 1. Probability of s�rvival in R �the level of ser�m antibodies above or eq�al to median M� a red line� vs WR �the level below M� a bl�e line�: a� b� c — the relation of the anti­TF­pAA IgG level to s�rvival; d — the relation of the anti­αGal IgG level to s�rvival. a — all patients; b — patients with an N��� stat�s; c� d — patients with G� t�mors Experimental Oncology ��� �������� ���� ���ne���� �������� ���� ���ne� ���ne� ��9 Table 2. The relation of the antibody level in the serum of patients to survival Anti- bodies Group n p HR Mean survival time, months and 95% CL* TF-pAA All 56 0.012 0.22 185.8 (163.2–208.4) vs 121.2 (84.9–157.4) Stages II–III 40 0.008 0.23 169.7 (136.8–202.6) vs 95.8 (56.7–134.9) T2–4 36 0.062 0.30 168.1 (130.0–206.2) vs 100.3 (53.3–147.3) N1–2 28 0.021 0.23 164.2 (123.1–205.3) vs 81.8 (38.1–125.6) G3 25 0.017 0.18 167.5 (122.1–212.9) vs 68.7 (26.8–110.5) Tn Stages II–III 42 0.075 2.49 108.3 (66.5–150.1) vs 157.9 (125.2–190.6) αGal All 59 0.101 2.43 147.0 (115.0–178.9) vs 181.3 (160.8–201.7) G3 27 0.066 3.27 98.4(40.4–156.3) vs 159.1 (121.1–197.2) Note: *R vs WR. 95% confidence limit (CL) is lower and upper confidence limits shown in brackets. Breast cancer patients that were described in re- ference [19] are also included in the Table ba Fig. 2. IEF and the IgG­Western blotting of samples affinity­ isolated from the ser�m of cancer patient: a — pAA­reactive antibodies isolated on TFα­spacer­Sepharose; b — pAA­reactive antibodies isolated on TFα­PAA­Sepharose. a and b correspond to samples � and � that described in [��]. The arrows show the difference in intensity of IgG­bands. The cathode on the top DISCUSSION We demonstrated earlier an association of the level of examined antibodies with s�rvival of patients with gastrointestinal cancer incl�ding pathomorphological s�bgro�ps. A significantly better s�rvival has been ob­ served in patients with an increased level of antibodies to TF­pAA and Tn­PAA while� on the contrary� a signifi­ cantly worse s�rvival was observed in an increased anti­ αGal­PAA IgG level or its postoperative elevation [��]. The present st�dy demonstrates an association of the ser�m level of anti­TF­pAA IgG and trend to associa­ tion of anti­Tn and anti­αGal IgG with s�rvival of breast cancer patients. Better s�rvival was revealed in an in­ creased level of anti­TF­pAA antibodies for all gro�p. Similar to patients with gastrointestinal cancer� a better s�rvival of TF­R was observed in s�bgro�ps of an ad­ vanced breast cancer and G� t�mor �see Table ��. Al­ tho�gh a significant differences for Tn­R vs Tn­WR and αGal­R vs WR were not fo�nd� val�es of HR>� and mean s�rvival time for s�bgro�ps presented in Ta­ ble � show a trend to worse s�rvival with an increased level of antibodies. The relation of anti­Tn and ­αGal antibodies level to s�rvival is less significant in breast cancer than in gastrointestinal cancer. The antibody response to some glycans of enteric microorganisms may be more informative for t�mors of digestive tract. A low level of preoperative anti­TF­pAA IgG domi­ nated in stages III�IV of gastric cancer or in metastases N���� as well as in poorly differentiated carcinoma G� and it was �nfavorable indicator of s�rvival [��� �9]. Noteworthy� TF­WR in s�bgro�ps N��� and G� de­ monstrated worse s�rvival either in gastrointestinal or in breast cancer. The lower level of anti­αGal IgG has been associated with an advanced breast cancer. A significantly lower anti­αGal IgG was noted for gre ater t�mor size �T� + T� vs T�� and for stage II vs I as well as for low­differentiated breast carcinomas G� vs G� + G� [�9]. However� patients with G� t�mor and a lower level of anti­αGal antibodies �αGal­WR� were prone to longer s�rvival �see Table ��. The low IgG reactivity to TF­pAA in patients with cancer may be d�e to the presence of ser�m factors that infl�ence the detection of antibodies. Nat�ral anti­glycan antibodies are mostly IgM­class antibo­ dies. IgM with corresponding specificities can compete with IgG for binding to glycan in ELISA and thereby red�ce IgG signal [��]. As a r�le� we observed a low IgM reactivity to TF­pAA in ser�m samples with a high corresponding IgG reactivity b�t inverse correlation was not revealed. No difference in s�rvival of gastric cancer patients was demonstrated in relation to the anti­TF­ pAA IgM level [��]. The other ser�m factor that can red�ce IgG binding to TF­pAA and thereby red�ce the signal of antibodies in ELISA is the TF­binding protein galectin �. Ser�m levels of galectin � in patients with breast and gastrointestinal cancer were significantly elevated especially in metastatic gastrointestinal carci­ noma as compared with healthy individ�als [��]. A high level of galectin � is associated with the worse s�rvival of colorectal cancer patients [��]. The increased level of galectin � in advanced cancer and its competition may be an additional factor improving eval�ation of s�rvival �sing determination of IgG reactivity �level� to TF­pAA. This s�pposition needs f�rther detailed investigations. Anti­glycan antibodies belong to nat�ral antibo dies. Comm�nities of microbiota transmitted from mother to newborn remain stable in lifetime. The nat�ral IgG antibodies are also transferred from mother to fet�s and the repertoire of IgG a�toantibodies remains stable [��� �4���]. We observed a similar pattern of anti­TF­pAA� ­Tn and ­αGal IgG in sera of grand­ mother� mother and her ad�lt son; moreover� everyone of donors had a high level of anti­TF­pAA and anti­αGal IgGs. In most gastrointestinal cancer patients the an­ tibodies level is slightly changed b�t in some patients ��� Experimental Oncology ��� �������� ���� ���ne� we observed a high level of anti­glycan IgG and its postoperative elevation [��� ��� ��]. A very high level of anti­TF­pAA IgG was marked in eight long­term s�rvivors in stages II�III among ��4 patients �nder st�dy [��]. The twelve­fold change of the anti­TF­pAA IgG level was noted in the follow­�p. In one case the stable increase over the backgro�nd level co�ld be d�e to prolonged chemotherapy. The anti­TF­pAA IgG level correlated with a co�nt of blood lymphocytes and in­ versely correlated with ne�trophil/lymphocyte ratio [��� ��]. Together these observations point to the adaptive anti­TF­pAA IgG response at least in some patients. Anti­glycan antibodies associated with s�rvival may be involved in cancer progression thro�gh different path­ ways. Some of them are cross­reactive to histo­blood gro�p related antigens that may be aberrantly expressed in t�mors as prec�rsors or incompatible antigens [��]. Some genera of microbiota might express TAG­mimic antigens and stim�late TAG­a�toreactive antibodies� which may be beneficial or adverse for cancer patients. To characterize the specificity of antibodies� anti­ TF­pAA IgGs were affinity­isolated from the ser�m samples of long­term s�rvivors� �sing different TFα­ conj�gated sorbents. Anti­TF­pAA IgG displayed a low specificity to the m�cino�s fraction isolated from malignant breast t�mors except three specimens [��� �9]. Affinity­isolated anti­Tn and ­αGal IgG also were low specific. The cross­reactivity of anti­TF­pAA IgG to different determinants incl�ding TFα­�nrelated reactivity and reactivity to the pAA carrier witho�t glycans was revealed. We renamed term �sed ear­ lier “anti­TF IgG” to “anti­TF­pAA IgG”. Antibodies were more reactive to TFβ �Galβ���GalNAcβ� than to TF �TFα� conj�gates� and they were a heterogenic pop�lation varying in the cross­reactivity to glycolipid­ related glycans with TFβ terminal resid�es� GA� and Gb5tri [��� ��� ��]� the latter being a terminal trisac­ charide of Gb5 �stage­specific embryonic antigen �� SSEA­��� which is expressed in ��.5% of breast cancer specimens [��]. The specificity to the pAA carrier and ��%­TFα­pAA b�t not to the standart ��%­TFα­PAA� which was inherent in all IgG s�bpop�lations incl�ding those isolated witho�t carrier� is enigma [��]. These IgG s�bpop�lations belong to polyclonal antibodies �see Fig. ��. The pAA­mimic �glyco�peptide seq�ences of �nknown proteins may be nat�ral determinants for antibodies. The identification of nat�ral targets co�ld el�cidate the direct or indirect relation of a long­lasting anti­TF­pAA IgG imm�ne response to s�rvival. Dysbiosis is associated with a breast cancer state and severity. The total bacterial DNA load was red�ced in the t�mor as compared to normal tiss�e and corre­ lated inversely with an advanced disease [�]. Dysbiosis may infl�ence the t�mor development via complex interrelations within the triad: resident microbiota — t�mor — imm�ne response in which some anti­glycan antibodies might be involved. The dynamic st�dy of the h�moral imm�ne response �sing glycomics together with microbial genomics can el�cidate interrelations leading to the carcinogenesis and t�mor progression. CONCLUSIONS The present st�dy demonstrates prognostic sig­ nificance of anti­TF­pAA IgG� which is acceptable not only for gastrointestinal cancer b�t also for breast can­ cer. The non­invasive screening test may be a �sef�l s�pplement in clinical o�tcome assessment. ACKNOWLEDGEMENTS This st�dy was s�pported by a grant № ��99 from the Estonian Science Fo�ndation. REFERENCES 1. Arrieta MC, Finlay BB. The commensal microbiota drives immune homeostasis. Front Immunol 2012; 3: 33. 2. Xuan C, Shamonki JM, Chung A, et al. Microbial dysbiosis is associated with human breast cancer. PLoS One 2014; 9: e83744. 3. Sheflin AM, Whitney AK, Weir TL. Cancer-promoting effects of microbial dysbiosis. Curr Oncol Rep 2014; 16: 406. 4. Stowell SR, Arthur CM, McBride R, et al. 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