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Immunological aspects of the juvenile respiratory papillomatosis of the larynx
Muhamadaminova Shoira Abduvalievna, Tashkent Medical Academy, ENT department, doctor-intern E-mail: Inf-ledi@mail.ru
Ismailova Adolat Abdurahimovna,
Nabieva Umida Pulatdjanovna
Immunological aspects of the juvenile respiratory papillomatosis of the larynx
Abstract: This study was based on the analysis of clinical and immunological observations of 36 children with respiratory laryngeal papillomatosis of the age from 7 months to 15 years. We managed to establish deep T-cellular immune deficit which expressed in inhibition of the number of T-lymphocytes and T-helpers/inductors on the basis of increase in T-cytotoxic lymphocytes, natural killers, serum immunoglobulin A and activation of IL-6, which is important prognostic criterion in the formation and development of tumors of viral origin.
Keywords: Juvenile respiratory papillomatosis, cellular and humoral factors of immune system, IL-6, prognostic criterion.
Background
The juvenile respiratory papillomatosis of the larynx (URP) is the most common benign tumor of the upper respiratory ways in children. Papillomatosis of the larynx accounts from 15.9 to 57.5 % of all benign tumors of the larynx. In the etiology of disease the intrauterine infection or contamination through the maternal passages have the main importance [1, 67-68; 4, 63-91; 8, 111-113; 19, 300].
Despite of significant increase of knowledge about etiology and immunopathogenesis of the laryngeal papillomatosis in the last decade many questions concerning mechanisms of the development of pathological process and progressing remain to be open. Until recently mechanisms of immune response were studied at a level of separate populations and cells of immune system, and such deeper mechanisms as cytokines, were insufficiently examined [6, 4-10; 7, 36-40; 14, 1645-1651; 15, 23]. It is known, that in development and progressing of the viral lesions the cellular and humoral factors of the immune system, particularly cytokines, have the important role, which are endogenous biologically active substances realizing intercellular interaction [7, 36-40]. Last years the more data have been collected about significance of immunological mechanisms in the development of one or another pathology иммунологических of mechanisms in development of this or that pathology, including at viral lesion of the larynx. Thus, according to the literature data infectious-inflammatory larynx processes result in deep damages of immunological reactivity [3, 8-12; 4, 63-91; 9, 37-40; 10, 22-25; 12, 777-786].
In this connection, we carried out evaluation of cellular and humoral factors of immune system as well as there was studied concentration of anti-inflammatory cytokine IL-6 and anti-viral interferonalpha in the serum of peripheral blood of children with juvenile respiratory papillomatosis of the larynx.
Material and methods
This study was based on the analysis of clinical and immunological observations of 36 children with respiratory laryngeal papillomatosis of the age from 7 months to 15 years receiving treatment in the ENT clinic of Tashkent Medical Academy (TMA). The period of follow-up was from 3 months to 10 years. This period of observation, on out opinion, is enough for discussion of efficacy of various methods of treatment and outcomes of disease.
There was isolated the group (out of total number) of patients with primary and frequent recurrent laryngeal papillomatosis
consisted of 14 and 22 children, respectively. Control group included 29 children of the analogue age and sex.
Immunological examination included determination of the
neutrophilephagocytary activity, phagocytary index, blood serum concentrations of the major immunoglobulins (IgG, IgA, IgM) and circulating immune complexes in the serum of the peripheral blood (CIC big and small). Cellular immune parameters were measured by the contents ofleucocytes, lymphocytes, total pool of T-lymphocytes (CD3+), T-helpers/inductors (CD4) and T-suppressors/cy-totoxic lymphocytes (CD8+), ratio CD4/CD8 (immune regulatory index — IRI), B-lymphocytes (CD20+), as well as by expression of activate markers (CD38+, CD95+) on the lymphocytes. Immu-nological examinations were performed in the laboratory of im-munocytokines of the Institute of Immunology of the Academy of Sciences of the Republic of Uzbekistan. The PCR diagnosis of YPV DNA in the brash-biopsies was performed, where there was identified type 11 VPC in all the studied patients. The serum concentrations of IL-6 and IFN-alpha were measured by IFA method with use of commercial test-systems "HUMAN" (Germany).
Results and discussions
In the laryngeal papillomatosis the content of leucocytes in the peripheral blood, on the average, was not differed reliably between each other and in comparison with control. In group of children with primary process the reliable increase in comparison with lymphocyte content was noted in the control group and group of children with frequent- recurrent papillomatosis. It is known that CD3+, CD4+, CD8+ receptors are related to phenotype markers ofT-lymphocytes. Analysis of the immunophenotype of T-lymphocytes in children with laryngeal papillomatosi showed reliable inhibition of CD3+ expression on the T-lymphocytes and its absolute value in group of children with primary papillomatosis in comparison with control group and children with frequent recurrences (P < 0.05). Evidently, reduction of the general pool of T-lymphocytes (CD3+) was found due to number of T-lymphocytes, expressing marker CD4+. It is known that CD4+ T-cellular response to viral proteins is important mechanism of microorganism defense, because CD4+ T-helpers stimulates production of the antibodies by B-lymphocytes and activated CD8+ T-lymphocyte, specific against virus-infected cells [3, 8-12; 14, 1645-1651]. Thus, in the groups of children with papillomatosis of the larynx there was observed reliable inhibition of the CD4+ expression on the lymphocyte in comparison with control group. Interesting, that the most inhibition of the relative number of T-helper/inductor was charac-
Section 6. Medical science
teristic for group of children with frequent recurrent papillomatosis (p < 0.05). In group of children with primary papillomatosis CD4+ T-helpers/inductors were reduced 1.5 times, and in the frequent recurrent — 1.8 times in comparison with control values. It is known, that cytotoxic CD8+ T-lymphocytes play the important role in he pathogenesis ofviral diseases [3, 8-12]. The function of these cells is recognition of antigens on the cellular surface in complex with molecules of class 1 MHC. CD8+ T-lymphocytes play the leading role in the virus elimination, that may be explained, on the one side, by their ability to induce death of the infected cells, expressing the appropriate peptides, presenting by molecules of class 1 MHC, and, on the other side, by the ability to secreate antiviral factors. (proinflammatory cytokines — IFN-a, TNF-a and others) [7; 9]. Thus, in group of children with frequent-recurrent papillomatosis there was noted reliable increase ofthe relative content of CD8+ T-lymphocytes in comparison with data of control group and group of children with primary papillomatosis of the larynx. Consequently, immuneregulatory index in group of children with frequent recurrent papillomatosis of the larynx was considerably inhibited and was 0.81 ± 0.02 (p < 0.05). Obviously, the decrease in immune regulatory index is an important criteria of the deepness of T-cellular immune deficit state in the frequent-recurrent papillomatosis of the larynx.
The natural killer cells (NKC), which are the third population of lymphocytes providing supporting of genetic homeostasis, which were differed significantly from T- and B-lymphocytes by phenotype and functions. They are related to the category of the main effectors of natural or congenital immunity, which are able to lyse cells-targets or to induce antibody dependent cellular toxicity. NKC participate in the antiviral, antibacterial and antiprotozoya defense. We revealed reliable increase in relative and absolute values CD16+ in the both studied groups of children in comparison with values of control group. It was shown that parallel with T-lymphocytes B-lymphocytes are the main effectors of immunity. The function of B-lymphocyte in struggle of body against infection is the production of antibodies. Thus, the relative and absolute values of B-lymphocytes in all studied children with laryngeal papillomatosis remained in the control limits that testify to active participation of B-lymphocytes in the antiviral immune response. And at the same time, it is known that their protective efficacy under the conditions ofviral persistence is limited [17, 27; 18, 144]. It was established that functional insufficiency of B-lymphocytes in the antiviral immune response was characterized by production of immunoglobulins [17]. They play important function as mediators in the cascade development of immune response and partially may be responsible for efficacy of final effector reactions of cellular immunity for inactivation and elimination ofviral antigens [5, 311-312; 17, 27]. The investigations showed that serum concentrations of the main IgG and IgM in children with papillomatosis of the larynx remain to be in the limits of the control indicators, and serum content of IgA was reliably increased in the both studied groups of children in comparison with control group (p < 0.05), however, there were no reliable differences between groups.
Study of activational lymphocyte markers particularly in infectious disease, especially in papillomatosis because analysis of activational lymphocyte markers allows to study processes of activation, proliferation, differentiation and apoptosis of immunocompetent cells and characterizes cellular cycles associated with processes is of great scientific and practical significance [5, 311-312]. CD8+ is the activational marker which is considered as multifactorial
protein that is ectoenzyme catalyzing synthesis and hydrolysis of CADF-ribose. Fermentative functions of CD38+ provide its main immuneregulatory role that is binding of various agents by
this receptor that provides enhancement of the cytokine synthesis, kinase activation and protein phosphorilation [5, 311-312]. CD38+ is the precursor of the plasmatic cells which expresses on the immature T- and B-lymphocytes, activated T-lymphocytes, plasmacytes. We revealed reliable increase in expression of CD38+ on the lymphocytes in the both studied groups of children with papillomatosis (p < 0.05). According to literature data there is evidence about role of APO-1/Fas (CD95+) receptors during apoptosis, and its degree reflects lymphocyte apoptosis level [5, 311-312]. The growth of the expression of receptor CD95+ on the lymphocytes indicated about apoptotic way of lymphocyte death. Thus, we showed increased expression of CD95+ on the lymphocytes of peripheral blood of children with laryngeal papillomatosis, and significant increase in apoptosis of the lymphocytes was revealed in the group of children with primary papillomatosis. Evidently, the excessive apoptosis in papillomatosis in combination with humoral immunity and deep T-cellular immune deficit contributes to progressing disease. It is known that CD23+ is specific receptor of the immunoglobulin E. This receptor presents on the surface of 30 % of B-lymphocytes and 1 % of the t-cells and monocytes. Under the influence of IL-4 CD23+ begins to be produced by B-cells and monocytes in the soluble form [5, 311-312; 7, 36-40]. It is known that high levels of CD23+ are observed in infectious diseases [5, 311-312]. Expression of the CD23+ molecule is rather increased in children with laryngeal papillomatosis, particularly in children with primary papillomatosis.
One of the important directions in the modern medicine working on the junction with molecular biology, is to study the role of cytokines in the pathogenesis of the infectious diseases and their complications. The cytokines are considered as proteins of low molecule weight which are produced by cells of various cells and are mediators of intercellular interactions in immune response [8]. We studied the serum concentration of proinflammatory cytokine IL-6 and antiviral interferon-alpha (IFN-a). Comparative analysis of IL-6 in children with papillomatosis of the larynx in various studied groups revealed presence of the reliable difference with values of control groups (p < 0.05). In group of children with frequent-recurrent papillomatosis there was found reliable increase in value of IL-6 in comparison with data of children with primary laryngeal papillomatosis. The data obtained indicated about important diagnostic role of IL-6 in progressing of laryngeal papillomatosis. It is known that IL-6 is glycoprotein that is pleiotropic cytokine of broad diapason of biological activity and is produced both by lymphoid and non-lymphoid cells of the organism. It was established that IL-6 regulates immune and acute phase response, inflammation, oncogenesis and hemopoiesis [5, 311-312; 7, 36-40]. One of the main functions of IL-6 is regulation of the processes of maturation of the antibody-producing cells from B-lymphocytes and of production itself of immunoglobulins. Thus, we revealed considerable increase in IL-6 serum concentration in the group of children with laryngeal papillomatosis, and maximal value of IL-6 was noted in group of children with frequent recurrent papillomatosis of the larynx.
It is established that disturbance of the cytokine balance, directed to hyperproduction of IL-6, is accompanied by excessive symptoms of inflammation and sometimes it is the central chain of the pathogenesis of viral lesions at the systemic level. It is known that asthenic syndrome was the characteristic clinical manifestation of the laryngeal papillomatosis in children, such signs of disease as weakness, flaccidity, subfebrile temperature, poor appetite which are under control of the cytokine system. Similar suggestions are supposed by other authors [6, 4-10; 7, 36-40; 9, 37-40; 17, 27; 18, 144].
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The system of interferons is the integral part of the immune system which provides coordination of proliferation, differentiation and activation of effectory cells of the immunity. The interferon status is the assessment of the functional state of the interferon system [7]. It is known that IFN-a, playing one of the main roles in the viral elimination, has direct antiviral effect as well as indirect immunomodulationg activity. IFN-a is produced almost by all the cells of the body, but most of all by macrophages and lymphocytes. IFN-a is the strong antiviral protein [8]. Analysis of the results showed that the serum content of IFN-a in the group of children with primary papillomatosis of the larynx was reliable increased in comparison with data of the group of children with frequent recurrences and children of control group. Thus, in group of children with primary recurrence the level of IFN-a was increased 3.5 times in comparison with control group
and 2.4 times — with values of children with frequent recurrences. Evidently in cases of frequent recurrences of disease the exhaustion of the reserves of the interferon system that is accompanied by inhibition of the IFN-a production.
Thus, our investigations allowed revealing reliable changes in the functioning of the immune system of the children with laryngeal papillomatosis. Our purpose was to find the most vulnerable indicators of the immunity which may be served in the further as important diagnostic, therapeutic and prognostic criteria in laryngeal papillomatosis in children. We managed to establish deep T-cellular immune deficit which expressed in inhibition of the number of T-lymphocytes and T-helpers/inductors on the basis of increase in T-cytotoxic lymphocytes, natural killers, serum IgA and activation of IL-6, which is important prognostic criterion in the formation and development of tumors ofviral origin.
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Makhmudova Aziza,
Research Scientific Institute of Hematology and Blood Transfusion, MoH of Uzbekistan Republic Uzbekistan, Tashkent E-mail: innaberger@mail.ru Karimov Khamid Yakubovich, Makhmudova Aziza Djumanovna, Shamsutdinova Dildora Bakhtiyarovna, Boboev Kodirjon Tuxtaboevich, Berger Inna Viktorovna
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