Aberrant and multiaberrant (rogue) cells in peripheral lymphocytes of Hodgkin's lymphoma patients after chemotherapy

We analyzed spontaneous chromosome lesions in peripheral lymphocytes cultured from Hodgkin's lymphoma (HL) patients before and after cytostatic chemotherapy. The mean aberration frequency was significantly higher in HL patients after chemotherapy (7.20+/-0.58 per 100 metaphases) than in non-treated HL patients (4.80+/-0.54), and in non-treated patients than in healthy subjects (2.12+/-0.13). In lymphocytes of HL patients, who received chemotherapy, we found, in addition to ordinary aberrant cells, a large number of multiaberrant (or rogue) cells, i.e. metaphases carrying multiple (at least four) chromosome-type exchange aberrations. Rogue cells were found in 15 out of 18 chemotherapeutically treated HL patients (in total, 60 rogue cells per 5,568 scored cells), whereas in 30 non-treated patients only 1 rogue cell was found (per 4,988 scored cells). No correlation was found between the yield of rogue cells and the aberration frequency in ordinary aberrant cells. Aberration spectra (ratios of chromatid- to chromosome-type aberrations and of breaks to exchanges) were essentially different in ordinary aberrant and multiaberrant cells. These data, as well as analysis of cellular distributions of aberrations, implied independent induction of chromosome damage in ordinary aberrant and rogue cells. Analysis of aberration patterns in diploid and polyploid rogue metaphases belonging to the first, second, and third in vitro division indicated that rogue cells could be formed both in vivo and in vitro, and could survive at least two rounds of in vitro replication, given blocked chromosome segregation. These results suggested that formation of rogue cells, unlike ordinary aberrant cells, was triggered by events other than direct DNA and/or chromosome lesions. A hypothesis regarding disrupted apoptosis as a candidate mechanism for rogue cell formation seems to be most suitable for interpretation of our data. Cultured lymphocytes of chemotherapeutically treated HL patients may represent a model system for further examination of the multiaberrancy phenomenon.     

Time-effect relationship of chromosome aberrations in peripheral lymphocytes after radiation therapy for seminoma

The time-effect relationship of dicentrics and cells containing unstable chromosome abnormalities (Cu cells) was studied in peripheral lymphocytes of 40 blood samples from 23 patients suffering from seminoma during a time period of 0-1720 days after radiation therapy. Nine patients were studied before treatment. Since the half-time for the disappearance of damaged cells from circulating blood is an increasing function of post-exposure time it can only be expressed as a differential value. The present model discriminates between the mean lifetime m for lymphocytes and a parameter q which is the differential half-time for the decline of damaged cells immediately after exposure (t = 0). If the time t is short compared with m the decline is asymptotically time-hyperbolic rather than exponential and can be described by q only. According to recalculations of previous data, comprising 30 years post exposure, m approximates 10 years. Differential half-times can be derived for any time post treatment within the analysed time period for the decline of the incidence of dicentrics. For example at the end of therapy (t = 0) the differential half-time was calculated to be 0.4 years and at 1720 days post exposure 3.6 years resulted. The corresponding values for the percentage of Cu cells cannot be derived for t = 0; at 1720 days 3.9 years resulted.     

Evaluation of the small bowel in patients with Hodgkin's lymphoma during and late after chemoradiation therapy

The paper describes the experience in using capsule endoscopy to diagnose small bowel lesions during and after chemoradiation therapy in patients with diagnosed Hodgkin's lymphoma and an attempt to compare the current views of normal tissue response to ionizing radiation, as well as drugs used for chemotherapy.     

Genotoxic effects of radiotherapy and chemotherapy on circulating lymphocytes in patients with Hodgkin's disease

Chemical mutagenesis of Caenorhabditis elegans has relied primarily on EMS to produce missense mutations. The drawback of EMS mutagenesis is that the molecular lesions are primarily G/C --> A/T transitions. ENU has been shown to produce a different spectrum of mutations, but its greater toxicity to C. elegans makes it a difficult mutagen to use. We describe here methods for minimizing ENU toxicity in C. elegans. Methods include preparing ENU stocks in absolute ethanol and storing stock solutions for not more than 2 weeks at -20 degrees C. To maintain reasonable brood sizes of mutagenized animals, mutagenic solutions should not exceed 1.0mM ENU. We provide data which suggest ENU is degraded or altered to more toxic products in aqueous solution, but less so in solvents such as absolute ethanol.     

The appearance of immature cell leukemias after radiologic and cytostatic intensive therapy of Hodgkin's lymphoma]

Among 106 patients with Hodgkin's lymphoma in stage IIIB and IVB treated from September 1969 to June 1977 by a working team for hematology and oncology at the clinic and outpatients clinic of the Medical School of Erfurt, there were two cases (=1.9%) showing a final transition to leukaemia with immature cells. A third neoplasia which had developed in a former field of irradiation was additionally observed in one patient. The various possibilities arising from an oncologic effect of ionizing rays and those substances having a cytostatic effect are discussed. After frequent reports on the occurrence of secondary tumours following intensive radiological and cytostatic therapy of advanced Hodgkin's lymphoma the enhanced risk of a secondary tumour being induced by this combined therapy cannot be excluded. Various conclusions are drawn from that by the authors.     

Micronuclei in lymphocytes of prostate cancer patients undergoing radiation therapy

To further verify the applicability of the micronucleus (MN) assay in biodosimetry, we measured the MN yield in cytokinesis-blocked (CB) peripheral blood lymphocytes (PBL) of eight prostate cancer (PC) patients. These patients had no previous chemotherapy or radiotherapy (xRT). They were treated with standardized schemes of fractionated pelvic xRT. Before xRT, and at one random time-point during the course of xRT, blood samples were collected from each patient for the following purposes: (1) to verify the relationship between the MN yield in PBL and the estimated equivalent (EQ) total-body absorbed dose; and (2) to evaluate the individual differences of ex vivo radiation dose-response (1-4 Gy) relationship of MN yield in PBL before xRT. The number of xRT fractions, cumulative tumor dose, and EQ total-body absorbed doses of these patients represented a wide range. We found in PBL of these patients that (1) MN yield (Y) increased linearly with the estimated EQ total-body absorbed dose as Y=14.6+9.2D (R(2)=0.7, p=0.007); the distributions of MN yield were overdispersed; the ratio of relative increment of MN yield per 1000 binucleated (BN) PBL ranged from 0.9 to 8.2 (median: 4.1) folds above that of the respective baseline levels; and (2) before xRT, the MN yields also increased linearly with the ex vivo radiation dose; at each radiation dose level, the distributions of MN yield were overdispersed in most patients. In two of the three patients with xRT-induced early side effects (cystitis, diarrhea), the MN yield in PBL induced by ex vivo irradiation before xRT was significantly higher than in the other patients without xRT-induced side effects. These findings suggest that MN yields in CB PBL can be used as an in vivo biodosimeter. Since the differences in individual ex vivo radiation dose-response relationship of MN yield in PBL before xRT appeared to be significant, our preliminary results also suggest that it may be possible to identify individual intrinsic radiosensitivity before the start of xRT.     

Frequency and distribution of mitomycin C-induced structural chromosome aberrations in lymphocytes from non-Hodgkin lymphoma patients

Lymphocytes from 15 untreated patients with non-Hodgkin lymphoma (NHL) and 15 controls were exposed to 0.08 micrograms/ml mitomycin C, and the frequency and distribution of structural chromosome aberrations (chromatid and chromosome gaps, breaks, and exchanges) were analyzed in 100 mitoses per subject. The mean frequencies of aberrant cells, and gap, break, and gap + break events were 8.7, 0.9, 9.7, and 10.6 in the NHL group and 11.6, 1.1, 12.7, and 13.8 in the control group. None of the differences between the two groups was significant (P greater than 0.05). The distribution of breakpoints was nonrandom (P less than 0.001) in both groups, with a particularly marked excess of breaks in 9q11. The other breakage-prone bands were 1q11 and 1q21 in the NHL group and 1p11, 1q11, 2q31, and 16q11 in the control group. None of these hot spots coincided with any of the 60 bands known to be involved in primary chromosome abnormalities in NHL.     

The chromosome aberrations in human peripheral blood lymphocytes 43-46 years after the acute radiation disease

Cytogenetic study of workers, who had an acute radiation syndrome of the medium (ARS II), severe (ARS III) and extremely severe (ARS III-IV) degrees in 1953-1957, was performed. Lymphocytes from peripheral blood were cultured and analyzed with using the routine chromosome staining (4 individuals) and FISH (2 individuals) methods. In each case 4000-1000 metaphase slides were analyzed with the group chromosome kariotyping. A high frequency of chromosome aberrations (CA) was revealed, i.e.: 9.33-9.8 CA per 100 cells for ARS II patients, 28.6 and 36.6 CA per 100 cells for patients with the severe ARS. The main type of rearrangement is stable CA (up to 90%). The CA frequency exceeds the level of spontaneous CA (control--20 individuals) and CA of the patients, who had Chronic Radiation Disease (CrRD) 45 years ago (20 individuals). By 43-46 years of the control. No cancer diseases or hematopoietic pathology were revealed by 43-46 years of follow-up.     

Molecular-biological properties of blood lymphocytes of Hodgkin's lymphoma patients. Plausible possibility of treatment effect prognosis

Hodgkin's lymphoma (HL) patients were investigated before and during chemical and radiation therapy. The properties of peripheral blood lymphocytes of the HL patients before treatment have been compared with healthy donors and the patients during the treatment. The genetic damage--frequency of cells with micronuclei (MN), the level of DNA single- and double-strand breaks (SSB and DSB), DNA-protein cross-links (DPC) have been studied. Biochemical and physiological parameters have been compared as well: the concentration of reactive oxygen species (ROS), the ability to the adaptive response induction. The radiosensitivity of lymphocytes in vitro exposed to the 1 Gy irradiation has also been determined (by MN test). It was shown that in Hodgkin's lymphoma patients' lymphocytes (in comparison with healthy donors) the frequency of cells with MN does not change, the level of SSBs and DSBs increases, the amount of DPC does not change, and ROS concentration (on average) significantly increases because of the part of the population that have high ROS content. The ROS concentration decreases to control level, the frequency of cells with MN increases, the level of DSBs does not change but the level of DPCs (which prevents the determination of DSB) increases in the patients during treatment. It was also discovered that lymphocyte radiosensitivity correlates with the MN cells frequency before treatment and the ROS concentration. These results make it possible to suppose that the high MN frequency and high ROS concentration in Hodgkin's lymphoma patient lymphocytes (before treatment) can serve as prognostic factors for the effectiveness of radio and chemical therapy.     

An analysis of the distribution and dose response of chromosome aberrations in human lymphocytes after in vitro exposure to137Cesium gamma radiation

The chromosome aberration yield for human lymphocytes exposed in vitro to various doses of 137Cesium has been studied. Dicentric, total acentric, and excess acentric data were seen to follow a Possion distribution. Calculated total hits demonstrated over-dispersion which could possibly be accounted for by a greater occurrence of single-hit phenomena being repaired than two-hit exchange processes. The resulting distribution generally contained an under-representation of cells with odd numbers of hits and an over-representation of zero- and even-hit classes as compared with Poisson predicted values. The relationship between dicentric yield and dose received in rads was fitted to the linear-quadratic formula Y = alpha D + beta D2 for dicentrics, yielding values of (20.1 +/- 3.8) X 10(-4) (aberrations/cell)/rad and (1.89 +/- 0.75) X 10(-6) (aberrations/cell)/rad2 for alpha and beta respectively. A plot of percent 'normal' cells versus the dose in rads resembled cell survival curves and was fitted to the relation P(D) = 100 e-Y where Y = alpha D + beta D2 with alpha = (23 +/- 11) X 10(-4) rad-1 and beta = (8.3 +/- 2.5) X 10(-6) rad-2. A possible use of scoring 'normal' cells for purposes of biological dosimetry is presented.     

Differences of arteria carotis in patients with Hodgkin's lymphoma

Radiotherapy increases the risk of cardiovascular morbidity. We examined arteria carotis atherosclerosis and stenosis in Hodgkin's lymphoma patients. We examined arteria carotis of 120 Hodgkin's lymphoma patients who have been in complete remission for at least 5 years. 70 patients received neck irradiation (mean age at the time of the examination was 44.6 years). Twenty-four (34.3%) of them had carotis sclerosis or stenosis, and it was significantly more than in the control group [8 out of 60 patients 13.3%)]. Twelve patients of the 50 who did not receive radiotherapy had carotis lesions, and there was no significant difference compared to the control group. Significant stenosis (>50%) was detected in only 3 patients (in the irradiated group). TIA, stroke or amaurosis fugax did not occur. Carotis stenosis does not seem to play a role in late mortality in Hodgkin's lymphoma, but if the patient has an increased risk for atherosclerotic changes, then regular examinations are necessary, and other risk factors (smoking, hypertension, diabetes mellitus, hypothyroidism, early menopause) need to be treated.     

The intrauterine origins of Hodgkin's lymphoma

Background: Both small and large body size at birth are now known to predict a range of chronic disorders in adult life, including certain cancers. These associations are thought to reflect “fetal programming.” This may lead to impairment of a small number of key systems including the immune system. Hodgkin's lymphoma is a disease of the immune system. We have therefore examined the association between Hodgkin's lymphoma and early development. Our hypothesis was that the disease would be associated with markers of poor fetal growth, specifically small body size or small placental size at birth. Methods: Using the Finnish Cancer Registry we identified patients with Hodgkin's disease in a cohort of 20,431 people born in Helsinki during 1924–44. Each person has a detailed birth record. Results: There were 12 patients with Hodgkin's disease, giving an incidence comparable to international rates. The disease was associated with prolonged gestation. For every additional week of gestation the hazard ratio was 1.37 (95% CI 1.00–1.87; p = 0.05). The disease was also associated with a short placental surface. After allowing for gestation, for each centimetre increase in surface length, the hazard ratio was 0.70 (0.53–0.92; p = 0.01). The disease was not associated with birth weight or length or maternal body size. Conclusions: We have shown that Hodgkin's lymphoma is associated with prolonged gestation and a short length of the placental surface. We speculate that Hodgkin's lymphoma is initiated by two events in fetal life. One, which is an immune event, is associated with prolonged gestation, while the other is associated with growth faltering.     

Plasma biomarkers for detecting Hodgkin's lymphoma in HIV patients

The lifespan of people with human immunodeficiency virus (HIV) infection has increased as a result of effective antiretroviral therapy, and the incidences of the AIDS-defining cancers, non-Hodgkin''s lymphoma and Kaposi sarcoma, have declined. Even so, HIV-infected individuals are now at greater risk of other cancers, including Hodgkin''s lymphoma (HL). To identify candidate biomarkers for the early detection of HL, we undertook an accurate mass and elution time tag proteomics analysis of individual plasma samples from either HIV-infected patients without HL (controls; n = 14) and from HIV-infected patient samples with HL (n = 22). This analysis identified 60 proteins that were statistically (p<0.05) altered and at least 1.5-fold different between the two groups. At least three of these proteins have previously been reported to be altered in the blood of HL patients that were not known to be HIV positive, suggesting that these markers may be broadly useful for detecting HL. Ingenuity Pathway Analysis software identified “inflammatory response” and “cancer” as the top two biological functions associated with these proteins. Overall, this study validated three plasma proteins as candidate biomarkers for detecting HL, and identified 57 novel candidate biomarkers that remain to be validated. The relationship of these novel candidate biomarkers with cancer and inflammation suggests that they are truly associated with HL and therefore may be useful for the early detection of this cancer in susceptible populations.     

The molecular mechanisms of classic Hodgkin's lymphoma

Classic Hodgkin's lymphoma is characterized by the appearance of giant abnormal cells called Hodgkin and Reed-Sternberg (HRS) cells. HRS cells arise from germinal center B lymphocytes and in about 50 percent of patients, are infected with Epstein-Barr Virus. In addition, HRS cells show constitutive NF-kappaB activation and are resistant to apoptosis. This paper reviews several recent studies that for the first time implicate specific molecules in the pathogenesis of classic Hodgkin's lymphoma. Targeting these molecules could lead to the development of novel therapies for this disease.     

Chromosome abnormalities in peripheral blood lymphocytes from untreated Hodgkin's patients

Summary We describe the presence of a high frequency of spontaneous chromosome aberrations in lymphocytes from six untreated patients with Hodgkin's disease. The characteristics of the chromosome abnormalities observed suggest the existence of a certain degree of chromosome instability in these cases, that could be a predisposing factor for the development of malignancies.     

Lipid peroxidation and protein oxidation in patients affected by Hodgkin's lymphoma

A dysregulation of the redox homoeostasis has been reported in various neoplastic disorders. Malondialdehyde/4-hydroxy-2,3-nonenal (MDA/HNE) and protein carbonyl groups represent in vivo indexes of lipid peroxidation and protein oxidation, respectively, suitable to investigate radical-mediated physio-pathological conditions. We evaluated MDA/HNE and protein carbonyl groups in sera of untreated Hodgkin's lymphoma (HL) patients in advanced disease stages, in order to quantify the oxidative stress. HL patients displayed significantly higher levels of both MDA/HNE and protein carbonyl groups as compared with healthy controls. This is the first evidence that a strong increase in HL is one of the most common haematological malignancies, representing approximately 30% of all lymphomas in the circulating protein carbonyl content in HL. These findings may contribute to a better definition of the redox homoeostasis dysregulation in HL.     

The characterization and transmissibility of chromosome aberrations induced in peripheral blood lymphocytes by in vitro alpha-particle radiation

Peripheral blood lymphocytes were irradiated in vitro with (213)Bi alpha particles at doses of 0, 10, 20, 50, 100, 200 and 500 mGy. Chromosome analysis was performed on 47-h cultures using single-color fluorescence in situ hybridization (FISH) to paint chromosomes 1, 3 and 5. The whole genome was analyzed for unstable aberrations to derive aberration frequencies and determine cell stability. The dose response for dicentrics was 33.60 +/- 0.47 x 10(-2) per Gy. A more detailed analysis revealed that the majority of aberrations scored as dicentrics were part of complex/multiple aberrations, with the proportion of cells containing complexes increasing with dose. Cells containing aberrations involving painted chromosomes (FISH aberrations) were further classified according to cell stability and complexity. The majority of cells with FISH aberrations were unstable. The proportion of aberrant FISH cells with complex/multiple aberrations ranged from 56% at 10 mGy to 89% at 500 mGy. A linear dose response for genomic frequencies of translocations in stable cells fitted the data from 0 to 200 mGy with a dose response of 7.90 +/- 0.98 x 10(-2) per Gy, thus indicating that they are likely to be observed in peripheral blood lymphocytes from individuals with past or chronic exposure to high-LET radiation. Comparisons with the dose response for low-LET radiation suggest an RBE of 13.6 for dicentrics in all cells and 3.2 for translocations in stable cells. Since stochastic effects of radiation are attributable to genetic changes in viable cells, translocations in stable cells may be a better measure when considering the comparative risks of different qualities of radiation.     

Regularities in forming chromosome aberrations in cattle lymphocytes in external and combined radiation exposure

Aberrations in lymphocytes of cattle blood exposed to gamma-radiation and combined radiation were found. It was shown that seven days after the exposure to doses from 64.5 to 103.3 mC/kg a number of aberration varied in the range from 16.1 to 37.7 per 100 cells, whereas a frequency of aberrated cells was from 13.5 to 29.8. After the exposure to 77.3 mC/h and the following inclusion of fused radioactive particles into the fodder (48.1-762.2 Mbq/kg of the live weight), the number of aberrations increased from 25.5 up to 55.0 per 100 cells and the number of aberrated cells increased from 21 up to 43%.