Sister Chromatid Exchange Rates Research Articles

Genomic instability in patients with Barrett's esophagus

Our study aimed to determine, by counting sister chromatid exchange (SCE) and micronucleus (MN) frequencies, whether genetic impairment and DNA damage have an effect on the pathogenesis of Barrett's esophagus (BE). This study was conducted between June 2007 and November 2008 in the Erzurum Training and Research Hospital. We analyzed SCE and MN frequencies in 30 patients with BE, and in 30 control cases. SCE was significantly increased in BE patients compared with controls (6.89 ± 1.04 vs. 5.01± 0.88, P < 0.001). Similarly, MN was significantly increased in BE patients compared with controls (3.48 ± 1.08 vs. 1.83 ± 0.64, P < 0.001). Our data indicate that the increased SCE and MN rates in lymphocytes of patients with BE may reflect genomic instability or deficiency of DNA repair capacity.

The role of ascorbic acid on titanium dioxide-induced genetic damage assessed by the comet assay and cytogenetic tests

Titanium dioxide (TiO 2) is used in several commercial products such as cosmetics, sunscreen, toothpaste and pharmaceuticals. However, some recent investigations have revealed that titanium particles generate potential harmful effects on the environment and humans. Because of its strong antioxidant activity, ascorbic acid (AA) is admitted to act as an anti-mutagenic agent. The present study was undertaken to investigate the protective effect of AA against TiO 2-induced genotoxicity. Sister chromatid exchange (SCE), micronucleus (MN) and the comet assays were used to assess TiO 2-induced genotoxicity and to establish the protective effects of AA. There were significant increases ( P<0.05) in both SCE and MN frequencies of cultures treated with TiO 2 as compared to controls. However, co-application of AA (4.87 and 9.73 μM) and TiO 2 resulted in decreases of SCE and MN rates as compared to the group treated with titanium alone. Besides, significant reductions of primary DNA damage (comet assay) were determined when the AA was added to the cell culture medium simultaneously with TiO 2. In conclusion, the preventive role of AA in alleviating TiO 2-induced DNA damage was indicated for the first time in the present study.

Time to Bloom

Bloom Syndrome (BS) is an autosomal recessive disorder due to mutation in Bloom helicase (referred in literature either as BLM helicase or BLM). Patients with BS are predisposed to almost all forms of cancer. BS patients are even today diagnosed in the clinics by hyper-recombination phenotype that is manifested by high rates of Sister Chromatid Exchange. The function of BLM as a helicase and its role during the regulation of homologous recombination (HR) is well characterized. However in the last few years the role of BLM as a DNA damage sensor has been revealed. For example, it has been demonstrated that BLM can stimulate the ATPase and chromatin remodeling activities of RAD54 in vitro. This indicates that BLM may increase the accessibility of the sensor proteins that recognize the lesion. Over the years evidence has accumulated that BLM is one of the earliest proteins that accumulates at the site of the lesion. Finally BLM also acts like a "molecular node" by integrating the upstream signals and acting as a bridge between the transducer and effector proteins (which again includes BLM itself), which in turn repair the DNA damage. Hence BLM seems to be a protein involved in multiple functions - all of which may together contribute to its reported role as a "caretaker tumor suppressor". In this review the recent literature documenting the upstream BLM functions has been elucidated and future directions indicated.

Elevated dNTP levels suppress hyper-recombination in Saccharomyces cerevisiae S-phase checkpoint mutants

MEC1, the essential yeast homolog of the human ATR/ATM genes, controls the S-phase checkpoint and prevents replication fork collapse at slow zones of DNA replication. The viability of hypomorphic mec1-21 is reduced in the rad52 mutant, defective in homologous recombination, suggesting that replication generates recombinogenic lesions. We previously observed a 6-, 10- and 30-fold higher rate of spontaneous sister chromatid exchange (SCE), heteroallelic recombination and translocations, respectively, in mec1-21 mutants compared to wild-type. Here we report that the hyper-recombination phenotype correlates with lower deoxyribonucleoside triphosphate (dNTP) levels, compared to wild-type. By introducing a dun1 mutation, thus eliminating inducible expression of ribonucleotide reductase in mec1-21, rates of spontaneous SCE increased 15-fold above wild-type. All the hyper-recombination phenotypes were reduced by SML1 deletions, which increase dNTP levels. Measurements of dNTP pools indicated that, compared to wild-type, there was a significant decrease in dNTP levels in mec1-21, dun1 and mec1-21 dun1, while the dNTP levels of mec1-21 sml1, mec1-21 dun1 sml1 and sml1 mutants were ∼2-fold higher. Interestingly, higher dNTP levels in mec1-21 dun1 sml1 correlate with ∼2-fold higher rate of spontaneous mutagenesis, compared to mec1-21 dun1. We suggest that higher dNTP levels in specific checkpoint mutants suppress the formation of recombinogenic lesions.

Cytogenetic effects of recombinant interferon-γ on lymphocytes cultures from patients with non–small cell lung cancer

Therapeutic effects of human interferons (IFN) on malignancies and infectious diseases have been demonstrated in several clinical trials. The effects of IFN alone or combined with other treatment modalities (radiotherapy and chemotherapy) in lung cancer are under investigation. Experimental data suggest that some cytokines, such as IFN-α and IFN-γ, exhibit cytogenetic properties in human normal lymphocytes from peripheral blood, but the mechanisms are not clear. The aim of the present study was to investigate the in vivo cytotoxic and cytostatic activity of IFN-γ. Patients with certain cases of non–small cell lung cancer not eligible for chemotherapy or chemoradiotherapy were treated with thoracic radiotherapy. After tumor relapse, local treatment with instillations of IFN-γ through the fiberoptic bronchoscope followed. To clarify the cytogenetic activity of IFN-γ, sister chromatid exchange (SCE) and proliferation rate index (PRI) were evaluated in lymphocyte cultures from these patients' peripheral blood samples immediately after diagnosis (baseline), 30 days after radiotherapy, and after the fifth instillation of IFN-γ. Our results show a decrease in SCE frequency and PRI values in lymphocytes after treatment with IFN-γ, suggesting that IFN-γ does not have cytotoxic activity but, in contrast, may induce repair mechanisms, as shown in earlier studies in other biologic models.

Cytoprotective activity of amifostine on cultured human lymphocytes exposed to irinotecan

Irinotecan (camptothecin, CAM) is a topoisomerase-I inhibitor with a well established action in the chemotherapy of colorectal and ovarian cancer. Hematological and intestinal toxicity are commonly noted in patients treated with CAM. In this study, we examined the cytoprotective efficacy of amifostine (ethyol, ETH) against chromosomal damage induced by this drug on cultured peripheral human lymphocytes. Cultured lymphocytes were exposed to CAM (50 and 100 ng/ml of final concentrations) without or with ETH (in concentrations varying between 40 and 800 μg/ml of final culture volume). CAM’s genotoxicity was quantified by counting the Sister Chromatid Exchange (SCEs) rate. The mitotic index (MI) and proliferation rate index (PRI) were also assessed. The SCE rate was increased following incubation with CAM, but the combined treatment of CAM with ETH significantly reduced the SCE formation, especially when ETH added at high concentrations. The MIs and PRIs remained also unaltered in cultures with CAM, but MIs were reduced with the combined treatment at high ETH concentrations. Clinical studies are required to assess the predicted benefits from ETH in patients receiving CAM.

Loss of Bloom syndrome protein destabilizes human gene cluster architecture

Bloom syndrome confers strong predisposition to malignancy in multiple tissue types. The Bloom syndrome patient (BLM) protein defective in the disease biochemically functions as a Holliday junction dissolvase and human cells lacking functional BLM show 10-fold elevated rates of sister chromatid exchange. Collectively, these phenomena suggest that dysregulated mitotic recombination drives the genomic instability underpinning the development of cancer in these individuals. Here we use physical analysis of the highly repeated, highly self-similar human ribosomal RNA gene clusters as sentinel biomarkers for dysregulated homologous recombination to demonstrate that loss of BLM protein function causes a striking increase in spontaneous molecular level genomic restructuring. Analysis of single-cell derived sub-clonal populations from wild-type human cell lines shows that gene cluster architecture is ordinarily very faithfully preserved under mitosis, but is so unstable in cell lines derived from BLMs as to make gene cluster architecture in different sub-clonal populations essentially unrecognizable one from another. Human cells defective in a different RecQ helicase, the WRN protein involved in the premature aging Werner syndrome, do not exhibit the gene cluster instability (GCI) phenotype, indicating that the BLM protein specifically, rather than RecQ helicases generally, holds back this recombination-mediated genomic instability. An ataxia-telangiectasia defective cell line also shows elevated rDNA GCI, although not to the extent of BLM defective cells. Genomic restructuring mediated by dysregulated recombination between the abundant low-copy repeats in the human genome may prove to be an important additional mechanism of genomic instability driving the initiation and progression of human cancer.

Effect of DNA Hypomethylation on Genotoxicity and Apoptogenicity of Sodium Arsenite in Laboratory Mice

The present study aims to evaluate the effect of DNA hypomethylation state on genotoxicity and apoptogenicity induced by sodium arsenite (NaAsO2) in normal adult male SWR/J mouse bone marrow cells. Animals were treated with intraperitoneally (i.p.) injected with (2.25, 4.50 or 9 mg kg(-1) b.wt. of NaAsO2 which represent 0.25, 0.50 or 1 of LD5, respectively) and killed 24 h later. Another different group of male mice was treated with three doses of 5-Azacitidine (5-AzaC), 5 mg kg(-1) b.wt. each dose and 3 h intervals between them. NaAsO2 administered after 6 days of the last dose. The three single doses of sodium arsenite alone significantly (p<0.05) increased the rate of total structural Chromosomal Aberrations (CAs), rate of Sister Chromatid Exchanges (SCEs), micronucleus (MNs) formation, PARP and Lamia-A degradation and apoptosis as compared with the negative control. The combined treatment with hypomethylation agent 5-AzaC significantly increased the rate of SCEs induced by NaAsO2 at low dose. Moreover, this treatment significantly increased the rate of polyploidy at all combined used doses. Furthermore, this treatment induced apoptosis at all used doses. The present study has shown that DNA hypomethylation had a negative effects represented in rate of (CAs), polyploidy, PARP degradation and apoptosis induced by (NaAsO2). On the other hand, DNA hypomethylation had positive effects represented in decreas rate of pulverized chromosomes, centromeric attenuations, (SCEs), (MNs) formation, prevent Lamina-A degradation and apoptosis.

Antigenotoxic, Antioxidant and Lymphocyte Induction Effects Produced by Pteropodine

Pteropodine is a heterohimbine-type oxindole alkaloid specifically isolated from 'Cat's claw' (Uncaria tomentosa), a plant that has shown cytostatic, anti-inflammatory and antimutagenic properties and is used in traditional medicine to cure a number of diseases. In this report, we studied the ability of pteropodine to decrease the rate of sister-chromatid exchanges and micronucleated polychromatic erythrocytes in mice administered doxorubicin. We also determined its capacity to induce lymphocyte production in mice as well as its free radical scavenging potential by applying the DPPH assay. We found pteropodine (100-600 mg/kg) to significantly decrease the frequency of sister-chromatid exchanges and micronucleated polychromatic erythrocytes in mice administered with 10 mg/kg of doxorubicin. Furthermore, we determined that pteropodine partially corrected bone marrow cytotoxicity induced by doxorubicin, as it showed an improvement in the rate of polychromatic erythrocytes. Besides, 600 mg/kg of pteropodine increased 25.8% of the production of lymphocytes over the control value along a 96-hr assay, and it exhibited a strong capacity to trap the DPPH-free radical (98.26% with 250 microg/ml). Our results establish that pteropodine is an effective antimutagen in the model used, and suggest that pteropodine deserves further research in the area of cell protective potential and its mechanism of action.

Longitudinal biomonitoring of nurses handling antineoplastic drugs

To assess a possible trend in the genotoxic risk of oncologic nurses during the working year, cytogenetic biomonitoring was performed. Exposure to cytostatic agents is a major occupational concern in oncologic personnel. In contrast to the controlled environment in oncology pharmacies, nurses may be subject to unexpected events of exposure due to the intensive contact with patients. The entire nursing staff of an oncology inpatient ward (n = 15) participated in a biomonitoring study over a period of nine months using the sister chromatid exchange test and the comet assay to detect DNA strand breaks. Blood samples were taken after a three-week summer break (base level), one, three, six and nine months thereafter. Airborne contaminations of cytotoxics were addressed by chromatographic methods. With regard to the single monitoring points, the comet assay revealed no significant alteration of the genotoxic burden within nine months. By contrast, the sister chromatid exchange levels were significantly increased after six and nine months when compared with base levels. A trend analysis covering the whole observation period revealed an increase in genotoxicity as shown by the sister chromatid exchange test and the alkaline but not the neutral comet assay. This increase, however, was small and reversible as shown by the trend analysis of sister chromatid exchange rates during the years of service. Air samples were negative for cytotoxic contaminants. The small, but statistically significant genotoxic burden observed in oncologic nurses of an inpatient ward emphasises the need for a continuing effort to eliminate residual occupational risks. In comparison with historical controls, the current situation is characterised by beneficial safety improvements over the last years. Nevertheless, periodic training and awareness of the problems should be an integral part of advanced education.

Role of Recombination in the Long-Term Retention of Transposable Elements in rRNA Gene Loci

Multiple theoretical studies have focused on the concerted evolution of the tandemly repeated rRNA genes of eukaryotes; however, these studies did not consider the transposable elements that interrupt the rRNA genes in many organisms. For example, in insects, R1 and R2 have been stable components of the rDNA locus for hundreds of millions of years, suggesting either that they have minimal effects on fitness or that they are unable to be eliminated. We constructed a simulation model of recombination and retrotransposition within the rDNA locus that addresses the population dynamics and fitness consequences associated with R1 and R2 insertions. The simulations suggest that even without R1 and R2 retrotransposition the frequent sister chromatid exchanges postulated from various empirical studies will, in combination with selection, generate rDNA loci that are much larger than those needed for transcription. These large loci enable the host to tolerate high levels of R1 and R2 insertions with little fitness consequences. Changes in retrotransposition rates are likely to be accommodated by adjustments in sister chromatid exchange (SCE) rate, rather than by direct selection on the number of uninserted rDNA units. These simulations suggest that the rDNA locus serves as an ideal niche for the long-term survival of transposable elements.

The Saccharomyces cerevisiae checkpoint genes RAD9, CHK1 and PDS1 are required for elevated homologous recombination in a mec1 (ATR) hypomorphic mutant

Specific ataxia telangiectasia and Rad3-related (ATR) mutations confer higher frequencies of homologous recombination. The genetic requirements for hyper-recombination in ATR mutants are unknown. MEC1, the essential yeast ATR/ATM homolog, controls S and G2 checkpoints and the DNA damage-inducibility of genes after radiation exposure. Since the mec1-D (null) mutant is defective in both S and G2 checkpoints, we measured spontaneous and DNA damage-associated sister chromatid exchange (SCE), homolog (heteroallelic) recombination, and homology-directed translocations in the mec1-21 hypomorphic mutant, which is defective in the S phase checkpoint but retains some G2 checkpoint function. We observed a sixfold, tenfold and 30-fold higher rate of spontaneous SCE, heteroallelic recombination, and translocations, respectively, in mec1-21 mutants compared to wild type. The mec1-21 hyper-recombination was partially reduced in rad9, pds1, and chk1 mutants, and abolished in rad52 mutants, suggesting the hyper-recombination results from RAD52-dependent recombination pathway(s) that require G2 checkpoint functions. The HU and UV sensitivities of mec1-21 rad9 and mec1-21 rad52 were synergistically increased, compared to the single mutants, indicating that mec1-21, rad52 and rad9 mutants are defective in independent pathways for HU and UV resistance. G2-arrested mec1-21 rad9 cells exhibit more UV resistance than non-synchronized cells, indicating that one function of RAD9 in conferring UV resistance in mec1-21 is by triggering G2 arrest. We suggest that checkpoint genes that function in the RAD9-mediated pathway are required for either homologous recombination or DNA damage resistance in the S phase checkpoint mutant mec1-21.

Increased formation of sister chromatid exchanges, but not of micronuclei, in anaesthetists exposed to low levels of sevoflurane

We have assessed, for the first time, genotoxicity (i.e. sister chromatid exchanges and micronuclei) in anaesthetists exposed to a single volatile anaesthetic (sevoflurane) without nitrous oxide. The anaesthetists were exposed to an 8-h time-weighted average of 0.2 parts per million sevoflurane. Internists served as non-exposed controls. Mean (SD) sister chromatid exchanges per cell were significantly higher in anaesthetists compared to internists (6.6 (0.9) vs 5.1 (0.8); p < 0.001) whereas median (IQR [range]) micronuclei per 1000 binucleated cells did not differ (9.5 (6.3-10.8 [2.0-15.5]) vs 8.5 (6.0-10.5 [3.0-25.5]), respectively). Although the anaesthetists were exposed to rather low concentrations of sevoflurane, this 30% increase of sister chromatid exchanges is in agreement with a recently reported 300% increase with a high level exposure to sevoflurane and nitrous oxide. Omitting nitrous oxide does not normalise increased rates of sister chromatid exchanges.

Effects of boric acid and borax on titanium dioxide genotoxicity

Titanium dioxide (TiO(2)) is a potential carcinogenic/mutagenic agent although it is used in many areas including medical industries and cosmetics. Boron (as boric acid and borax) has also well-described biological effects and therapeutic benefits. In a previous study, sister-chromatid exchanges (SCEs) and micronuclei (MN) rates were assessed in control and TiO(2)-treated (1, 2, 3, 5, 7.5 and 10 microm) human whole blood cultures. The results showed that the rates of SCE (at 2, 3, 5, 7.5 and 10 microm) and MN (at 5, 7.5 and 10 microm) formation in peripheral lymphocytes were increased significantly by TiO(2) compared with the controls. The present study also investigated the genetic effects of boric acid and borax (2.5, 5 and 10 microm) on cultures with and without TiO(2) addition. No significant increase in SCE and MN frequencies were observed at all concentrations of boron compounds. However, TiO(2)-induced SCE and MN could be reduced significantly by the presence of boric acid and borax. In conclusion, this study indicated for the first time that boric acid and borax led to an increased resistance of DNA to damage induced by TiO(2).

Three NewBLMGene Mutations Associated with Bloom Syndrome

Bloom's syndrome (BS) is a rare autosomal recessive disease predisposing patients to all types of cancers affecting the general population. BS cells display a high level of genetic instability, including a 10-fold increase in the rate of sister chromatid exchanges, currently the only objective criterion for BS diagnosis. We have developed a method for screening the BLM gene for mutations based on direct genomic DNA sequencing. A questionnaire based on clinical information, cytogenetic features, and family history was addressed to physicians prescribing BS genetic screening, with the aim of confirming or guiding diagnosis. We report here four BLM gene mutations, three of which have not been described before. Three of the mutations are frameshift mutations, and the fourth is a nonsense mutation. All these mutations introduce a stop codon, and may therefore be considered to have deleterious biological effect. This approach should make it possible to identify new mutations and to correlate them with clinical information.

Effects of phytoestrogen genistein on cytogenetic biomarkers in postmenopausal women: 1 year randomized, placebo-controlled study

To evaluate in a twelve-month, randomized placebo-controlled study whether pure administration of phytoestrogen genistein (54 mg/day) might reduce cytogenetic biomarkers in peripheral lymphocytes of postmenopausal women. A total of 57 postmenopausal women met the criteria and were randomly assigned to receive phytoestrogen genistein (n = 30) or placebo (n = 27). There was no significant difference in age, length of time since menopause or body mass index between the two groups. After one year, plasma genistein level was 0.14 ± 0.01 μmol/L in the control group and 0.72 ± 0.08 μmol/L in the genistein group (P < 0.0001). At baseline, sister chromatid exchange rate was 4.97 ± 2.17 in the control group and 4.96 ± 1.83 in the genistein group (P = 0.89). After one year, sister chromatid exchange rate was 4.96 ± 2.16 in the control group and 3.98 ± 1.14 in the genistein group (P < 0.05). High frequency cells count was 3% in the genistein group and 5% in the control group (P < 0.05) at the end of the study. Chromosomal aberration frequency was 5.55% in the control group at time 0 and 5.75% in the genistein group; after one year, the figures were 5.86% in the control group and 4.5% in the genistein group (P < 0.05). After one year, there was a negative relationship between sister chromatid exchange rate and plasma levels (r = − 0.43; P < 0.05) in the genistein group.Phytoestrogen genistein has been shown in postmenopausal women to be effective in the reduction of cytogenetic biomarkers. The protective effect on genoma damage appears to be a particularly promising tool in reducing the risk of cancer.

Sister chromatid exchanges and chromosome aberrations in von Recklinghausen's disease--a family study.

Five members of a family with von Recklinghausen's disease (v.R.d.) were studied with respect to the occurrence of sister chromatid exchanges (SCE) and chromosomal aberrations. The frequency of gaps, breaks and other chromosomal aberrations in cells from the v.R.d. family did not differ from those in controls. In the controls, the frequency of aberrations increased three-fold after treatment with mitomycin C, but in the v.R.d. family no response was observed. The frequency of spontaneous SCE was significantly lower in all but one of the members of the v.R.d. family than in the controls. Mitornycin C increased the rate of SCE in all the v.R.d. family members and also in the controls, the increase being small, however, in two members of the v.R.d. family. The low frequency of SCE may be specific to the disease.

Sister chromatid exchange analysis in patients with psoriasis

Psoriasis is a common, chronic inflammatory skin disease with unknown aetiology. An increased reactive oxygen species (ROS) and insufficient antioxidant activity have been determined in psoriatic lesions. The analysis of sister chromatid exchange (SCE) is a cytogenetic technique used to show DNA damage caused by an exchange of DNA fragments between sister chromatids. The study aimed to determine the rates of SCE in psoriatic patients (17 female and 19 male) and healthy controls (15 female and 15 male) as well as superoxide dismutase (SOD), glutathione peroxidase (GP) and catalase (CAT) activity in both groups. We found significantly higher SCE rates in the patients (P < 0.00001). In addition, statistically significant decreased levels of erythrocyte SOD and CAT activities were noted in the patients(P < 0.001 and P < 0.05 respectively). Furthermore, a statistically significant increased erythrocyte GP activity was found in the psoriasis group (P < 0.05). Our results indicate that chromosomal instability may play an important part in the aetiology of psoriasis. In addition, the results support the hypothesis of an imbalance in the oxidant-antioxidant system in psoriasis.

Telomere exchange and asymmetric segregation of chromosomes can account for the unlimited proliferative potential of ALT cell populations

Telomerase-negative cancer cells show increased telomere sister chromatid exchange (T-SCE) rates, a phenomenon that has been associated with an alternative lengthening of telomeres (ALT) mechanism for maintaining telomeres in this subset of cancers. Here we examine whether or not T-SCE can maintain telomeres in human cells using a combinatorial model capable of describing how telomere lengths evolve over time. Our results show that random T-SCE is unlikely to be the mechanism of telomere maintenance of ALT human cells, but that increased T-SCE rates combined with a recently proposed novel mechanism of non-random segregation of chromosomes with long telomeres preferentially into the same daughter cell during cell division can stabilize chromosome ends in ALT cancers. At the end we discuss a possible experiment that can validate the findings of this study.

Telomeric Recombination in Lymphocytes Implicates ALT, an Alternative Mechanism for Telomere Length Maintenance, in Normal Human Hematopoietic Cells.

Telomeres are the very ends of chromosomes and protect the genome from recombination, end-to-end-fusion, and recognition as damaged DNA. Telomeres are eroded with each cell division, eventually reaching such critically short length as to cause cell cycle arrest, apoptosis, or genomic instability. In most highly proliferative cells, including hematopoietic stem cells and T lymphocytes, telomere attrition is countered by telomere extension by telomerase reverse transcriptase complex. The majority of cancer cells also express telomerase, which maintains telomere length and allows indefinite cell proliferation. However, about 10% of tumors maintain telomere length in the absence of telomerase by mechanisms collectively termed alternative lengthening of telomeres (ALT). ALT mainly acts through asymmetrical exchange of telomeric material between chromosomes or sister chromatids, producing one daughter-cell with short telomeres and a limited life-span and its sister with long telomeres and higher proliferative capacity. To date, ALT has only been reported in cancer cells or through genetic engineering of mammalian cells. Here we investigated whether ALT mechanisms were active in hematopoietic cells using chromosome orientation fluorescent in situ hybridization (CO-FISH). In standard FISH, a telomeric probe produces fours signals per chromosome, one at each end of the two chromatids. Using CO-FISH, the newly synthesized DNA strand is fragmented by BrdU incorporation and UV light exposure and then digested by exonucleases. In CO-FISH, a telomeric probe produces two signals only, one at each end of the chromosome; in the presence of telomeric recombination, the telomeric signal is split, generating more than two signals per chromosome. Peripheral blood lymphocytes from three healthy volunteers, normal human fibroblasts, K562 cells, telomerase-positive HeLa cells (known to be negative for ALT),and telomerase-negative VA13 cells (known to be positive for ALT) were investigated for telomeric sister chromatid exchange (t-SCE); at least 20 metaphases per cell type were examined. Cultured peripheral blood lymphocytes and VA13 cells both showed increased levels of telomeric sister chromatid exchange in comparison to the other cells (P=0.0001): telomeric probe generated 2.62±0.11 telomeric signals/chromosome in lymphocytes; 2.23±0.04 in VA13 cells; 2.09±0.01 in HeLa cells; 2.02±0.01 in K562 cells; and 2.02±0.01 in human skin fibroblasts. Staining incorporated-BrdU over 24 hours and evaluation of “harlequin” chromosomes point to a similar rate of genomic sister chromatid exchange in lymphocytes, VA13 cells, and HeLa cells, suggesting that high chromatid exchange is confined to the telomeric region. A physical association between promyelocytic leukemia protein (PML) and telomeres is characteristic of some ALT-positive cells, but confocal microscopy failed to co-localize the telomeric probe and anti-PML monoclonal antibody in peripheral blood lymphocytes, suggesting that t-SCE in lymphocytes is not mediated by PML. This is the first demonstration of ALT activation in normal mammalian cells. ALT may be activated in peripheral blood lymphocytes as a complementary mechanism to maintain telomere length, and may explain the differences in age-related telomere shortening observed between lymphocytes and granulocytes.