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Siberian Medical University, Tomsk, Russia
Laboratory of Oncovirology, Institute of Oncology, Tomsk, Russia
Laboratory of Immunogenetics, Leiden University, The Netherlands
Эта статья опубликована в сборнике по материалы первой Международной юбилейной конференции «Актуальные проблемы инфектологии и паразитологии», посвященной 110-летию со дня открытия проф. К.Н.Виноградовым сибирской двуустки у человека (2-5 апреля 2001, г. Томск)
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РЕЗЮМЕ
Ранее проведенные исследования позволили установить, что при описторхозе у человека существенно повышено число клеток с цитогенетическими нарушениями. Имеются литературные и наши данные показывающие, что при описторхозе повышены титры антител к капсидам некоторых герпесвирусов. Кроме того в районах с наибольшей пораженностью описторхозом у местного населения наблюдается циркуляция одного из герпесвирусов - вируса Эпштеин-Барр. В этой работе мы задались целью установить зависимость между повышением уровня клеток с цитогенетическими нарушениями у больных описторхозом с активацией в организме этих больных вируса Эпштеин-Барр. Всего обследовано около 1500 человек, проживающих в двух регионах Сибири: 1) вблизи от бассеина реки Оби (здесь пораженность описторхозом была максимальной) и 2) вблизи от бассеина реки Енисея ( здесь пораженность описторхозом была черезвычайно редкой). Исследования показали, что наибольший уровень цитогенетических нарушений и самые высокие титры антител к капсиду вируса Эпштеин-Барр отмечаются в крови у жителей проживающих в бассеине реки Оби. Кроме того установлено, что у инвазированных описторхозом людей увеличен уровен IgG и IgM. Полученные нами данные подтверждают идею о том, что наблюдаемое повышение уровня клеток с цитогенетическими нарушениями у больных описторхозом может быть связано с активной персистенцией в организме этих людей вируса Эпштейна-Барр
ABSTRACT
Previous studies have shown an increase in the frequency of chromosomal abnormalities in the peripheral blood lymphocytes from opisthorchiasis patients. Some evidence suggests that there is an association between chronic opisthorchiasis and certain herpesviruses. To study the relationship of opisthorchiasis to the reactivation of Epstein-Barr virus (EBV) infection as well as the influence of opisthorchis infection on some cytogenetical and immunological parameters, we used the indirect immunofluorescence for measuring some virus specific antibodies, the cytokinesis-block lymphocyte micronucleus assay, and the quantitative immunodiffusion method for measuring immunoglobulin concentrations in serum. A total of 1,580 people were monitored in two comparable Siberian regions: in the Ob River region which is endemically related to opisthorchiasis caused by Opisthorchis felineus and in the nonendemic control Yenisey River region. There was no significant difference in each of the tested parameters between two uninfected controls from the endemic Ob and nonendemic Yenisey regions. We have found significant difference (p<0.01) in the frequency of micronucleated cytokinesis-block lymphocytes and the antibody levels against certain EBV antigens between the examined inhabitants of the opisthorchis-infected Ob and uninfected Yenisey regions. Furthermore, there were a good correlation (r +0.72) between the increase in titres of antibody to the EBV capsid antigen and the high frequency of micronucleated lymphocytes in the opisthorchis-infected Ob population. Also, levels of both IgG and IgM were increased in opisthorchiasis patients. This study confirms an association between chronic opisthrochiasis and reactivation of EBV which may be implicated in the development of cancer in opisthorchiasis patients.
INTRODUCTION
In 1884 Rivolta (Italy) [1] obtained from the liver of a cat and described a new species of the trematodes : Opisthorchis felineus. During the 1890s, the first cases of this helminthiasis were found among humans in many countries of Europe (Germany, Italy, France) and Russia, mainly in West Siberia within the Ob River region [2]. In 1904 Rindfleisch (Germany) reported on 40 opisthorchiasis patients and described the first liver cancer cases in such patients [3].
Among helminths, Opisthorchis felineus fits into a particular group, the trematodes. The developmental cycle of O. felineus was described by H. Vogel in 1934 [4]. These organisms undergo an alteration of generations with sexual reproduction taking place in the lumen of intrahepatic gall ducts of the definitive host (man or carnivorous animals) and asexual reproduction in the two intermediate hosts (snails and freshwater fish) [5].
The definitive host is infected by metacercariae of O. felineus through the consumption of raw or inadequately cooked freshwater fish. After getting into the duodenum the metacercaria loses its external envelope and migrates to the intrahepatic gall ducts. In the liver, parasites grow for over a month into adult hermaphroditic worms, which can live in the body of the definitive host for 25 years. The acute stage of opisthorchiasis is accompanied by eosinophilia, hepatomegaly, cholangitis, urticaria, and, often, diarrhea. The chronic stages of opisthorchiasis are characterized by chronic cholangitis and cholecystitis and may be complicated by cholangiocarcinoma [6,7]. The spread of O. felineus is limited to certain European and Siberian river regions. While the prevalence of opisthorchiasis among the population ranges from 1-10% in the Upper Ob to 85-100%, mostly among the autochthones, in the Lower Ob in West Siberia, there has never been an autochthonous case reported in the Yenisey River in East Siberia [2]. The Ob River region including the Ob River, its three large tributaries, namely the Tobol River, the Tom River, and the Irtysh River, and the numerous small tributaries is the largest endemic area of opisthorchasis caused by O. felineus in the world. Approximately 96% cyprinids in the Ob and Tom Rivers are affected by O. felineus [2]. In contrast, East Siberia has never been an endemic area because of total absence of snails B. inflata and B. troscheli (the first intermediate host) in the Yenisey River.
Between 1982 and 1986 chronic opisthorchiasis patients from several different areas within the Ob River region were cytogenetically examined for opisthorchis infection-related mutagenic effects using the chromosome analysis and the micronucleus test [8]. In that study, the opisthorchiasis patients had significant increases of chromosomal aberrations and micronucleated lymphocytes compared with uninfected control persons. Therefore, these findings demonstrated that chronic opisthorchiasis is characterized by chromosomal abnormalities in man.
According to Ivanskikh and Bliznyuk [9] opisthorchis infection is associated with the reactivation of certain latent herpesviruses implicated in the development of human cancers and chromosomal damage.
In addition, the population in the Ob River region were shown to have markedly elevated levels of antibodies to the Epstein-Barr virus (EBV) capsid antigen [10]. On the basis of these data, it was hypothesized that the cytogenetic abnormalities in the opisthorchiasis patients are mainly caused by the reactivation of endogenous herpetic infection, especially EBV infection which is involved in the genesis of nasopharyngeal carcinoma, Burkitt’s lymphoma, and some other human cancers [11, 12]. To test this hypothesis, the relationship of chronic opisthorchiasis to the reactivation of EBV infection was investigated cytogenetically and serologically in comparable population groups of two Siberian regions. In addition, the influence of opisthorchis infection on the concentrations of all five classes of immunoglobulins in man was studied by measuring their concentrations in the sera of these groups.
MATERIALS AND METHODS
Study populations
During the winters of 1995-96 and 1996-97, a total of 1,580 people were examined by cytogenetical, serological, and immunochemical methods in 9 towns and villages situated in two Siberian regions (Table 1). Of these, 1,186 persons live in the endemic Ob River region including Ob, Irtysh, and Tom areas and 393 persons live in the nonendemic Yenisey River region. Mean levels of people harbored O. felineus in the endemic Ob region vary between 6 studied places from 21.4% in the village of Dolon to 87.1% in the town of Salekhard. Patient diagnosis was confirmed with both serologic test (enzyme-linked immunosorbent assay) and stool analysis for opisthorchiasis [13]. All the opisthorchiasis patients in the Ob group were found to have ova of O. felineus in their stools on at least one examination during the six-month period before this study. Microscopical stool examination of the uninfected controls from the endemic Ob River (220 persons) and nonendemic Yenisey River regions (393 persons) was negative for opisthorchiasis. Immunologic tests for opisthorchiasis were also negative in these groups.
Informed consent was obtained from each person prior to beginning this study. In each case, a individual examined had to fill out a questionnarie, by means of which we grouped people according to place of residence, place of birth, sex, smoking habits, year of birth, an amnesis of diseases, the presence of children with abnormalities in the family, still-born children, cancer diseases, X-ray examinations taken, and length of residence in the locality.
Each of the persons also described their diet in their questionnaires. In this study the qualitative and quantitative evaluation of each diet was based on the method of Giovannucci et al. [14]. For each of 60 food and beverage items listed a commonly used unit or portion size was specified and participants were asked how often, on average, over the past year they had consumed that amount of each food. Participants chose from among nine possible responses, which ranged from ‘never’ to ‘six or more times per day’. We also enquired about the brand, duration and frequency of multivitamin and individual vitamin supplement used and the types of fat commonly used. The questionnaire also contained an open-ended section for foods that were not listed. We computed nutrient intakes by multiplying the consumption frequency of each unit of food by the nutrient levels of the specified portions. Since vitamin A has an antimutagenic effect and can decrease radiation-related chromosome damage, we calculated vitamin A intake among examined individuals. We calculated retinol (performed vitamin A) intake as two third of the vitamin A activity (in IU) of dairy foods plus the total vitamin A activity in other animal products. Both infected and control population groups included in the study ranged in age from 5 to 40 years and were distributed in three approximately equal age subgroups, namely the 5-10 year-old children, the 11-16 year-old children, and the adult age group. The male-female ratio in both infected and control groups was one to one.
Cytogenetic method
Micronuclei are known to be formed from the lagging fragments or whole chromosomes during cell division. To make quantitative comparisons, it is important to know the frequency of cells (with and without micronuclei) that have undergone one division following exposure. This has become possible by blocking cytokinesis by cytochalasin B, which leads to binucleated cells.
Heparinized peripheral blood samples were obtained from each blood donor. Cytokinesis-block lymphocyte micronucleus assay was performed according to recommendations of Fenech and Morley [15]. Whole blood cultures were set up according to our standard experimental protocol [7,16]. Briefly, whole blood (0.5ml) was added to 5 ml of RPMI-1640 tissue culture medium (Sigma, USA) containing 15 % fetal calf serum and antibiotics. Proliferation was intiated by adding 0.15 ml phytohemagglutinin (PHA-M; Difco, USA). The cultures were incubated at 37 0C in a 5% CO2 atmosphere for 52 h. Cytochalasin B (Merck) was added to the cultures to block cytokinesis 24 h before harvesting. The microscopic slides were stained with Giemsa for detection of micronuclei in cytokinesis blocked lymphocytes. The frequency of micronucleated cytokinesis-block lymphocytes were evaluated in 1,000 cells per individual by two independent cytogeneticists using coded slides.
Serological method
Serum. Venous blood was obtained from an antecubital vein with a disposable syringe after the skin was cleaned with alcohol. The blood was allowed to clot at ambient temperature (22 0C) for 4-6 h and centrifuged at 1,000 g; the serum was transferred to another tube, capped, and stored at -20 0C until used.
Determination of antibodies to EBV. In addition to the cytogenetic analysis, we also determined the following indices in the peripheral blood donors: first, the level of serum G-antibody to the EBV capsid antigen; second, the level of virus specific A-antibody to the EBV capsid antigen; third, the level of specific G-antibody to the EBV early antigen complex. To measure these virus specific antibodies, we used the standard indirect immunofluorescence according to Henle and Henle [17].
Immunochemical method
Quantitative immunodiffusion plates. The influence of opisthorchis infection on the concentrations of all five classes of immunoglobulins in man was studied by measuring their concentrations in the sera of infected and control groups according to Mancini et al. [18]. Quantitative immunodiffusion plates were obtained from DiagnoTekh, Moscow, Russia; the agar contained sufficient antibody to specific immunoglobulins to form precipitation rings that were directly measurable. There were 24 holes in each plate. Standards for IgG, IgM, IgA, and IgD were obtained from DiagnoTekh. Standards were diluted to 50% and 25 % of the original concentration with 0.145 M NaCl and stored at -20 °C.
Quantitative immunoglobulin (IgG, IgM, IgA, IgD) determination. These were done using the DiagnoTekh immunodiffusion plates and standards at the temperature, time, and directions given by DiagnoTekh. Standards (100%, 50%, 25 %) were determined in duplicate on each plate. The wells were filled with 3 ml of serum by disposable capillary micropipettes. After brief immersion in acetic acid ( 5 % ) , the precipitation rings on the plates were measured with the aid of an eyepiece with a graduated reticle and indirect lighting on a black background. Unknowns on each plate were quantitated against a standard curve on that plate. The sera for all groups were diluted two- fold with 0.145 M NaCl before addition to the wells.
Quantitative IgE determination. These were performed by a radioimmunoassay [19] on a randomly selected group of the samples obtained from each age group. Examinations for the helminth eggs were performed both qualitatively and quantitatively [2, 20].
Statistical analysis
Statistical analyses of the data were made by Student’s ‘t’ test and correlation assay. All calculations were carried out using the SAS Release 6.04 software package. The statistical inferences were based on the level of significance P<0.01 and P<0.05.
RESULT
Figure 1 summarizes the data of percentages of the people who harbor O. felineus, who have micronuclei in their lymphocytes, and/or who show the reactivation of EBV infection among the studied populations of 9 places located in the endemic Ob and nonendemic Yenisey regions. While the percentages of the people suffered from opisthorchiasis in the Ob region vary between 6 studied places from 21.4 to 87.1%, there has never been a autochthonous case in the Yenisey region. A few positive serologic tests for opisthorchiasis were seen in the Atamanovo group; however, since subsequent careful examination of stool did not expose eggs of O. felineus, these tests were assumed to be falsely positive. The negative stool examinations for ova are supported by a continuing failure to find a bona-fide case of opisthorchiasis in the Yenisey population.
It was found that people who showed an increase in the frequency of micronucleated lymphocytes and/or titres antibody to EBV antigens in the Ob region were statistically significantly higher than in the Yenisey region. Among the Ob populations, especially high level of people with micronucleated lymphocytes were observed in the villages of Samus and Dolon which were exposed to radiation from the Siberian Chemical Complex (Tomsk-7) and the Semipalatinsk Atomic Test Site respectively during the 1950s.
When the levels of antiviral antibodies against EBV was analysed in different age groups of the infected Ob population we found an age-dependent increase in the G-antibody titres to EBV capsid antigen (Table 2).
Analysis of the results from the examined inhabitants of the two regions has shown a significant difference (p<0.01) in the frequency of micronucleated cytokinesis-block lymphocytes between the opisthorchis-infected Ob group and both uninfected control Yenisey and Ob groups. We have found no significant difference in the frequency of micronucleated cells and the other tested parameters between both control opisthorchis-uninfected Ob and Yenisey groups.
In the infected Ob population, the levels of micronucleated lymphocytes were two- to three-fold higher than those of the uninfected Ob and Yenisey populations (6.4±0.7% and 2.8±0.5% respectively). When the frequency of micronucleated lymphocytes was analysed in relation to age we found a good positive age-dependent correlation in the infected Ob population (Table 2). However, no significant difference (p>0.05) between males and females in this population was observed with a frequency of micronucleated lymphocytes (Table 3). Also, there was no significant difference between a group of individuals whose relatives were suffered from cancer and a group without cancer cases in their families. Nevertheless, smoking males with cancer cases in their families showed the highest micronucleated lymphocyte frequency (8.2 ±0.7% ). Among nonsmoking males without cancer patients in their families this index was 5.1±0.6% (p<0.01). We found no correlation between the presence of abnormal and still-born children, X-ray examinations, and the high frequency of micronucleated lymphocytes in the examined populations. In the radiation-exposed villages of Dolon and Samus, the migrants who moved to these villages after 1960 as well as their children born after 1960 showed a marked decrease (4.1±0.6% ) in the levels of micronucleated lymphocytes compared with the natives (7.7±0.9% ).
We separately evaluated small (less than 3 mm) and large micronucleus frequencies in the cytokinesis block lymphocytes in the Yenisey and Ob populations. The ratio of small micronuclei to large micronuclei in the lymphocytes was 1:1 in all the infected Ob and uninfected Yenisey populations excepting the radiation-exposed Samus and Dolon populations which showed about 92.3% small micronuclei. Deficiency of folic acid, vitamin B12, carotenes is known to lead to genomic instability in man. In this study, both the Ob and Yenisey groups showed no correlation between the level of vitamin B12 ( r -0.18; P>0.05), carotenes (r 0.33; P>0.05), or folic acid (r 0.27; P>0.05) consumption and the frequency of chromosome damage. Both the control Yenisey and Ob groups were approximately equal in consumption of these vitamins and the other food and beverage items.
All the opisthorchiasis patients in the infected Ob population showed positive serologic tests for EBV infection. The average antiviral antibody titres to the EBV capsid antigen and the EBV early antigen complex in the infected Ob population were significantly higher than in both uninfected control Yenisey and Ob groups ( Table 4).
Analysis of the results (Table 5) showed that there was a good correlation between the frequency of micronucleated cytokinesis-block lymphocytes and the high levels of G-antibody titres to EBV capsid antigen (r +0.72) in the opisthorchis-infected Ob population. The EBV-positive opisthorchiasis patients with considerably elevated G-antibody titres to EBV capsid antigen (more than 1:640) showed the highest mean level of micronucleated lymphocytes (14.8±1.0%). In addition, the mean levels of micronucleated lymphocytes for the EBV positive opisthorchis-infected groups with high G-anibody titres (from 1:160 to 1:320 and more than 1:640) were significantly different (p<0.01) from the EBV positive healthy donors of the opisthorchiasis-infected Yenisey region (14.8*1.0% and 3.9*0.7% respectively for titres more than 1:640) and there was no correlation between a frequency of micronucleated lymphocytes and increased G-antibody titres to EBV capsid antigen in the opisthorchis-uninfected Yenisey population.
Analysis of the concentrations of all five classes of immunoglobulins in the sera of the comparable populations showed that the levels of IgG were significantly higher in the opisthorchis-infected Ob population than in the control Yenisey population for all three age groups (Table 6).
The mean concentration of IgM in serum was significantly higher in the 11-16-year-old children of the infected Ob population than in those in the uninfected Yenisey population; the mean level of the serum IgM of the 5-10-year-old children from the infected Ob or the uninfected Yenisey populations did not differ significantly (Table 6), but the mean IgM levels of children in the Samus and Dolon groups were significantly lower than those in the other Ob populations as well as in the control Yenisey population.
The mean level of IgA in the adults was nearly twice that of the younger age groups but there was no difference between the opisthorchis-infected Ob and uninfected Yenisey populations in the 5-10 year or adult age groups. The mean level of IgA in the 11-16-year-old children of the Ob population was lower than that of the Yenisey population (Table 6). The mean level of IgD and IgE was similar in all three age groups of the Ob and Yenisey populations (Table 6).
DISCUSSION
The similarities in geographic location and radioactive contamination-related history of the Ob and Yenisey River regions, coupled with the endemic opisthorchiasis in the Ob region and total absence of opisthorchiasis in the Yenisey region, provided a unique opportunity to study a number of cytogenetic, immunologic and serologic parameters relative to this disease process.
The findings of our previous investigation [2, 7, 8] indicated that the majority of opisthorchiasis patients had significant increases in chromosomal aberrations and micronucleated lymphocytes as well as a depression of cellular response and a decrease in T-lymphocyte levels compared with uninfected control persons. It was suggested that immunosupression among the opisthorchiasis patients could be due to some factors produced by the parasite. A factor isolated from O. felineus is shown to can induce analogous lymphocyte-function impairment in experimentally treated mice and guinea pigs. In addition, immunosupression in the opisthorchiasis patients may contribute to the reactivation of certain latent herpesviruses, especially EBV infection, relative to chromosomal abnormalities in man [8, 21].
In the present study, the levels of micronucleated lymphocytes and/or antibody titres to EBV infection among the studied populations in the endemic Ob region are significantly higher than those in the nonendemic Yenisey region. Also, we found a good correlation between elevated antibody levels against certain EBV antigens and an increase in the frequency of micronucleated lymphocytes among opisthorchiasis patients. Micronuclei are known to be formed from the lagging fragments or whole chromosome during cell division. According to Rowe et al. [22], Ohshima et al. [23] EBV can integrate into the human genome and induce various chromosomal abnormalities which can give micronuclei.
Nevertheless, among Ob populations, especially high levels of people with micronucleated lymphocytes were observed in the villages of Dolon and Samus which are in close proximity to two main sources of radioactive contamination in Siberia, the Semipalatinsk Atomic Test Site and Siberian Chemical Complex (Tomsk-7) respectively. It was suggested that the cytogenetic effects in the inhabitants of these villages are mainly related to the worst routine and accidental releases of radiation from Siberian Chemical Complex and the Semipalatinsk Atomic Test Site during the 1950s. This suggestion may be also confirmed by a high percentage of small micronuclei in lymphocytes as well as a decrease in the level of micronucleated lymphocytes among the migrants who moved to these villages after 1960 and their children born after 1960 compared with the natives.
However, in the control Yenisey population, the mean levels of people with micronucleated lymphocytes in the village of Atamanovo, which is also located in the radioactive contaminated area close to Krasnoyarsk Mining Chemical Complex (Krasnoyarsk-26), are only slightly higher than in the other places of the Yenisey region but are significantly lower than in each of the places in the Ob region.
Estimation of dietary intake of vitamin B12, folic acid, and carotenes derived from self-reported dietary questionnaires have not shown any considerable differences between the Ob and Yenisey regions. We have not found any correlation between the frequency of chromosome damage and the levels of these vitamin consumption in both regions. Slight difference in nutrient intake between these two regions seems not to affect any of the tested parameters.
Elevated levels of immunoglobulins have been demonstrated in opisthorchiasis infections of men and experimental animals. Current investigations [2] are defining this increase relative to specific immunoglobulin classes (IgG, IgM, IgA, IgD, and IgE). Levels of both IgG and IgM increase in human opisthorchiasis [2]. Most (95 %) of the increase in IgG appears to be nonspecific and unrelated to specific opisthorchis antigens [2]. The data in the present report confirm that opisthorchiasis is associated with an increase in IgG, a change that is most striking in children (11-16 years).
In addition, the G-antibody titres to the EBV capsid antigen of the opisthorchis-infected Ob populations were significantly higher than those of the control Yenisey population. Therefore, the elevated levels of immunoglobulins G in the opisthorchiasis patients appear to may be related to reactivation of EBV.
The mean IgM level in the infected Ob children (11-16 years) was elevated over that of the comparable group of the Yenisey population, but there is no difference between the Ob and Yenisey populations in the other two age groups. The increased mean level in the 11-16-year-old Ob population may reflect the early response to the initial phases of the opisthorchis infection. The IgM level of the infected Samus and noninfected Dolon groups is significantly lower than that of the other Ob and Yenisey populations. Presumably this reflects a lower incidence of infectious disease. In addition, this may be associated with radioactive contamination of the areas from the Siberian Chemical Complex and the Semipalatinsk Atomic Test Site [24].
The lower mean level of IgA in the 11-16-year-old Ob group compared with the control Yenisey groups has no adequate explanation. There are no consistently significant differences in the mean IgD and IgE levels of the Ob and Yenisey groups in any age group.
In conclusion, this study confirms an association between chronic opisthrochiasis and reactivation of EBV infection which may be implicated as a cocarcinogenic and comutagenic factor in the development of cancer and cytogenetic effects in opisthorchiasis patients.
|
Table 1.
Studied places and populations in endemic Ob River region and the control Yenisey River region
|
| Groups |
Number of persons |
Age (in
years) |
Sex |
| |
|
5-10
|
11-16
|
17-40 |
male |
female |
| I.Opisthorchis
infected populations from the endemic Ob River region: |
966 |
322 |
322 |
322 |
483 |
483 |
| The Ob
area: |
|
|
|
|
|
|
| 1. the town of Salekhard |
158 |
52 |
53 |
53 |
79 |
79 |
| 2. the town of
Khanty-Mansyisk |
164 |
55 |
54 |
55 |
82 |
82 |
| 3. the town of Surgut |
127 |
42 |
42 |
43 |
64 |
63 |
| The
Irtysh area: |
|
|
|
|
|
|
| 4. the town of Tobolsk |
108 |
36 |
36 |
36 |
54 |
54 |
| 5. the village of Dolon |
179 |
60 |
60 |
59 |
89 |
90 |
| The Tom
area: |
|
|
|
|
|
|
| 6. the village of Samus |
230 |
77 |
77 |
76 |
115 |
115 |
| II. Uninfected
controls from the nonendemic Yenisey River region : |
393 |
131 |
131 |
132 |
196 |
197 |
| 1. the village of Atamanovo |
128 |
42 |
43 |
43 |
64 |
64 |
| 2. the town of Turukhansk |
120 |
41 |
40 |
40 |
59 |
60 |
| 3 .the town of Dudinka |
145 |
48 |
48 |
49 |
73 |
72 |
| III. Uninfected
controls from the endemic Ob River region
: |
220 |
72 |
73 |
75 |
111 |
109 |
| 1. The
Ob area |
106 |
33 |
37 |
36 |
51 |
55 |
| 2. The
Irtysh area |
114 |
36 |
38 |
40 |
56 |
58 |
|
Table 2.
Mean frequencies of micronucleated cytokinesis-block lymphocytes in the peripheral blood and mean levels of ova of Opisthorchis felineus in stools as well as the average G-antibody titres to Epstein-Barr virus capsid antigen in serum of three age groups of the opisthorchis-infected Ob population and the uninfected control Yenisey population. Values represent mean ±SE in this table
|
| Measure |
Uninfected
Yenisey population (control) |
Opisthorchis-infected
Ob population |
| |
Children |
Adult |
Children |
Adult |
| |
5-10 years |
11-16 years |
17-40 years |
5-10 years |
11-16 years |
17-40 years |
| |
n= 131 |
n=131 |
n=132 |
n=322 |
n=322 |
n=322 |
| The frequencies of micronucleated lymphocytes (%) |
1.3±0.3 |
1.4±0.5 |
1.6±0.5 |
4.3±0.7 b |
4.9±0.9 a |
7.8±1.1 a |
| The frequencies of ova O. felineus in stools (abs./mg) |
0 |
0 |
0 |
2.1±0.8 |
3.3±0.9 |
7.2±1.2 |
| Average IgG titres to the Epstein-Barr virus capsid antigen |
20.2±3.6 |
24.8±4.8 |
38.8±5.7 |
43.1±2.6
a |
63.3±4.9
a |
126.0±8.9
a |
Statistical levels: a P<0.01; b P<0.05
|
Table 3.
Mean frequencies of micronucleated cytokinesis-block lymphocytes in the peripheral blood of different groups of the opisthorchis-infected Ob River population and uninfected Yenisey control population (in %). Values represent mean ±SE in this table.
|
| Characteristics |
Uninfected Yenisey
population (control) |
Number
of individuals (n) |
Opisthorchis-infected Ob
population |
Number of individuals (n) |
| All |
2.8±0.5 |
393 |
6.4±0.7 a |
966 |
| Males |
3.1±0.6 |
196 |
5.8±0.6 a |
483 |
| Females |
2.6±0.5 |
197 |
6.6±0.9 a |
483 |
| Smoking
individuals |
3.2±0.7 |
276 |
6.9±0.8 a |
394 |
| Nonsmoking
individuals |
2.5±0.6 |
117 |
5.4±0.7 b |
572 |
| Smoking males with cancer cases in their family |
3.4±0.6 |
68 |
8.2±0.7 a |
103 |
| Nonsmoking males without cancer cases in their family |
2.6±0.5 |
46 |
5.1±0.6 b |
206 |
| Natives of the radiation exposed villages |
3.6±0.6 |
82 |
7.7±0.9 b |
193 |
| Migrants to the radiation exposed villages |
3.1±0.5 |
46 |
4.1±0.6 |
213 |
Statistical levels: a P<0.01; b P<0.05
|
Table 4.
The average antiviral titres to the Epstein-Barr virus capsid antigen and the Epstein-Barr virus early antigen complex in the opisthorchis-infected Ob populations and the control Yenisey population. Values represent mean ±SE in this table.
|
| Measures |
Uninfected
Yenisey population (control)
n=393
|
Opisthorchis-infected
Ob population
n=966
|
| Average IgA titres to the Epstein-Barr virus capsid antigen |
0.2±0.1 |
6.0±0.8
a |
| Average IgG titres to the Epstein-Barr virus capsid antigen |
32.1±9.7 |
98.3±11.2
a |
| Average IgG titres to the Epstein-Barr virus early antigen complex |
0.1±0.1 |
6.6±0.8
a |
Statistical levels: a P<0.01
|
Table 5.
Mean levels of micronucleated cytokinesis-block lymphocytes in the peripheral blood of the opisthorchis-infected Ob population and the uninfected control Yenisey population according to G-antibody titres to EBV capsid antigen. Values represent mean ±SE in this table.
|
| Titres IgG to the Epstein-Barr virus capsid antigen |
The
frequencies of micronucleated lymphocytes in the
peripheral blood |
| |
Healthy donors (control) |
Opisthorchiasis patients |
| |
n=393 |
n=966 |
| >1:40 |
1.5±0.4 |
1.7±0.4 |
| 1:160-1:1:320 |
2.4±0.5 |
8.2±0.7
a |
| >1:640 |
3.9±0.7 |
14.8±1.0
a |
Statistical levels: a P<0.01
|
Table 6.
Levels of immunoglobulins (IgA, IgG, IgM, IgD, IgE) in serum of three age groups of the opisthorchis-infected Ob population and the uninfected control Yenisey population. Values represent mean ±SE in this table.
|
| Measure |
Uninfected Yenisey
population (control) |
Opisthorchis-infected
Ob population |
| |
Children |
Adult |
Children |
Adult |
| |
5-10 years |
11-16 years |
17-40 years |
5-10 years |
11-16 years |
17-40 years |
| |
n= 131 |
n=131 |
n=132 |
n=322 |
n=322 |
n=322 |
| IgG in serum (mg/ml) |
12.4±1.3 |
12.3±1.1 |
16.1±1.5 |
16.0±1.2
a |
15.3±1.0
a |
19.1±1.5
a |
| IgE in serum |
0.002± 0.0005 |
0.003±0.0006 |
0.002±0.0004 |
0.003±0.0004 |
0.004±0.0003 |
0.003±0.0004 |
| (mg/ml) |
| IgM in serum (mg/ml) |
2.0±0.4 |
1.2±0.3 |
2.3±0.5 |
1.7±0.5 |
1.8±0.4 a |
2.4±0.6 |
| IgA in serum (mg/ml) |
1.2±0.3 |
1.2±0.5 |
2.6±0.7 |
1.1±0.4 |
1.3±0.4 |
2.4±0.5 |
| IgD in serum (mg/ml) |
0.3±0.2 |
0.25±0.4 |
0.2±0.1 |
0.35±0.2 |
0.3±0.2 |
0.2±0.1 |
Statistical levels: a P<0.01
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