Cutaneous B-Cell Lymphoblastic Lymphoma.

B-cell lymphoblastic lymphoma (B-LBL) is a malignant neoplasm of immature B cells that accounts for only 10% of all cases of lymphoblastic lymphoma. Most commonly, B-LBL presents as bony lesions, but in rare cases, the disease manifests cutaneously. We present a case of simultaneous cutaneous and systemic presentation of B-LBL in an otherwise healthy 28-year-old man in which the lymphoblastic infiltrate stained positive for CD79a, Tdt, CD10, and CD20. A diagnosis of cutaneous B-LBL was made, and systemic work-up revealed widespread involvement of the skin, bone, and lymph nodes. Review of all currently described cases of cutaneous B-LBL with or without systemic involvement revealed that the most frequently positive tumor markers were CD79a (92.3%), Tdt (90.6%), and CD10 (83.3%). Systemic involvement of B-LBL was found in nearly half of all cases with cutaneous presentation.

[Two Resected Cases of Primary Pulmonary Mucosa-Associated Lymphoid Tissue Lymphoma, Differently Diagnosed in Chest Tomography Scan].

Case 1: A 81-year-old man was admitted to our hospital because of a mass shadow on chest x-ray examination. Chest computed tomography (CT) showed a 1.5 cm nodule in the middle lobe of the right lung. We suspected a primary lung cancer and performed video-assisted right middle lobectomy. Histopathological examination showed a white, elastic, hard and solid 30 × 10 × 10 mm nodule with infiltration of small-to-medium-sized lymphocytes that were positive for CD20 and CD79a, and negative for CD10 and Cyclin D1 in immunohistochemical staining. We diagnosed mucosa-associated lymphoid tissue (MALT) lymphoma. Case 2: A 67-year-old woman was admitted to our hospital because of a mass shadow in the right upper lobe on chest x-ray and chest CT. As the lesion had not grow in 1 year, the patient strongly wanted it resected, therefore we performed wedge resetion of the right upper lobe via video-assisted thoracic surgery. Histopathological examination showed a white, elastic, hard and solid 25 × 25 × 16 mm nodule with infiltration of small-to-medium-sized lymphocytes that had positive staining of CD20 and CD79a, and negative staining of CD10 and Cyclin D1. We diagnosed MALT lymphoma. Primary lung MALT lymphoma shows a variety of shadows on chest CT, similar to lung cancer and other inflammatory diseases. Local therapies such as surgery and radiation therapy are effective against early stage MALT lymphoma, but there is no consensus of a standard surgery.

Diffuse Large B Cell Lymphoma Coexistent with Follicular Cell Lymphoma.

BACKGROUND: Diffuse large B cell lymphoma (DLBCL) and follicular cell lymphoma (FL) synchronously occurring in the same individual is unusual. The authors describe a rare case of DLBCL coexisting with FL at diagnosis in a male patient, with intestinal and lymph node involvement. METHODS: Hematologic investigation, intestinal tumor biopsy, bone marrow examination, cytogenetic analysis, and PCR were performed. RESULTS: The patient received three courses of R-CHOP regimen chemotherapy, the mesentery lymph nodes were reduced to 2.7 x 1.4 cm compared to 5.4 x 5.0 cm at diagnosis. However, the mesentery lymph nodes were increased to 4.7 x 3.5 cm after four additional courses R-CHOP treatment. Unfortunately, the patient refused chemotherapy treatment or hematopoietic stem cell transplantation (HSCT) and was lost to follow-up. CONCLUSIONS: The presence of DLBCL coexisting with FL at diagnosis is rare and has generally been the subject of isolated case reports. Further studies are required to better understand the biological insights of the disease and to propose an optimal management strategy for this type of lymphoma.

Functionalized multiwalled carbon nanotubes as ultrasound contrast agents.

Ultrasonography is a fundamental diagnostic imaging tool in everyday clinical practice. Here, we are unique in describing the use of functionalized multiwalled carbon nanotubes (MWCNTs) as hyperechogenic material, suggesting their potential application as ultrasound contrast agents. Initially, we carried out a thorough investigation to assess the echogenic property of the nanotubes in vitro. We demonstrated their long-lasting ultrasound contrast properties. We also showed that ultrasound signal of functionalized MWCNTs is higher than graphene oxide, pristine MWCNTs, and functionalized single-walled CNTs. Qualitatively, the ultrasound signal of CNTs was equal to that of sulfur hexafluoride (SonoVue), a commercially available contrast agent. Then, we found that MWCNTs were highly echogenic in liver and heart through ex vivo experiments using pig as an animal model. In contrast to the majority of ultrasound contrast agents, we observed in a phantom bladder that the tubes can be visualized within a wide variety of frequencies (i.e., 5.5-10 MHz) and 12.5 MHz using tissue harmonic imaging modality. Finally, we demonstrated in vivo in the pig bladder that MWCNTs can be observed at low frequencies, which are appropriate for abdominal organs. Importantly, we did not report any toxicity of CNTs after 7 d from the injection by animal autopsy, organ histology and immunostaining, blood count, and chemical profile. Our results reveal the enormous potential of CNTs as ultrasound contrast agents, giving support for their future applications as theranostic nanoparticles, combining diagnostic and therapeutic modalities.

[Chronic lymphocytic leukaemia presenting with specific cutaneous infiltrates in the absence of lymphocytosis: Two cases]. / Leucémie lymphoïde chronique révélée par des infiltrats cutanés spécifiques en l'absence d'hyperlymphocytose : deux observations.

BACKGROUND: Specific cutaneous infiltrates of chronic lymphocytic leukaemia (CLL) are rare. They occur after a mean disease duration of 3 years. CLL skin infiltrates as the primary manifestation of the disease have been reported, but a normal lymphocyte count at diagnosis is rare. We present two cases of CLL initially presenting in the skin, without lymphocytosis. PATIENTS AND METHODS: A 53-year-old man developed papulonodular lesions of the face and infiltrated plaques of the scalp, and an 85-year-old woman presented erythematous nodules of the face and neck. Histopathology revealed a lymphocytic infiltrate, consisting of small mature B-cells CD20+, CD79+, with an aberrant phenotype CD5+. CD23 was positive in one case. The two patients had no lymphocytosis, but immunophenotyping was characteristic of CLL. In the second case, there was a sub-mental adenopathy, histologic analysis of which was consistent with CLL. The CLL was classified as Binet stage A in the two cases. No disease progression was noted at follow-up. DISCUSSION: The unusual feature of these cases is the lack of lymphocytosis at diagnosis. Thus, the skin lesions resulted in further evaluations for CLL, although the diagnosis was not suggested by the blood count. CONCLUSION: Skin involvement in CLL does not appear to be a poor prognostic indicator, arguing in favour of recruitment of circulating monoclonal B-cells rather than an additional tumour mass.

A new score including CD43 and CD180: Increased diagnostic value for atypical chronic lymphocytic leukemia.

Moreau score has been used to differentiate chronic lymphocytic leukemia (CLL) from other mature B-cell neoplasms. However, it showed limitations in Asian patients. Therefore, we conducted a new score system replacing CD5 and CD23 with CD43 and CD180 to evaluate its diagnostic value of CLL. 237 untreated samples diagnosed with mature B-cell neoplasms were collected and were randomly divided into an exploratory and a validation cohort by a 2:1 ratio. The expression of CD5, CD19, CD20, CD23, CD43, CD79b, CD180, CD200, FMC7, and surface immunoglobulin (SmIg) were analyzed among all the samples. A proposed score was developed based on the logistic regression model. The sensitivity and specificity of the proposed score were calculated by ROC curves. CD43/CD180, CD200, FMC7, and CD79b were included in our new CLL score, which showed a sensitivity of 91.8% and a specificity of 83.1%. These results were confirmed in a validation cohort with a sensitivity of 90.5% (p = 0.808) and a specificity of 79.5% (p = 0.639). In CD5 negative or CD23 negative CLL group, the new CLL score displayed improved sensitivity of 79.4% compared to Moreau score and CLLflow score (41.2% and 47.1%, respectively). In atypical CLL group, the new CLL score showed improved sensitivity of 84.2% compared to Moreau score and CLLflow score (61.4% and 64.9%, respectively). This proposed atypical CLL score helped to offer an accurate differentiation of CLL from non-CLL together with morphological and molecular methods, particularly in Chinese patients with atypical immunophenotype.

BOB.1, CD79a and cyclin E are the most appropriate markers to discriminate classical Hodgkin's lymphoma from primary mediastinal large B-cell lymphoma.

AIMS: To clarify which immunohistochemical markers could be helpful in distinguishing between classical Hodgkin's lymphoma (cHL) and primary mediastinal B-cell lymphoma (PMBCL) to more narrowly define 'B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and cHL'. METHODS AND RESULTS: Two hundred and 83 cHLs and 51 PMBCLs were analysed on validated tissue microarray platforms with antibodies to BOB.1, CD15, CD20, CD23, CD30, CD79a, cyclin E, LMP-1, MUM1p, p63 and Oct2. The marker cut-off scores were calculated using receiver-operating characteristic curves. Markers with the highest positive predictive value for cHL were: CD15, cyclin E, LMP-1 (all 100%), MUM1p (93%) and CD30 (83%). High sensitivity was achieved only by CD30 (92%) and cyclin E (79%). Nineteen percent of PMBCLs were also positive for CD30, which led to a lower specificity of CD30 as regards cHL (81%) compared with cyclin E (100%). The antibodies with the highest positive predictive value for PMBCL were: CD23 (98%), p63 (96%), BOB.1 (94%) and CD79a (90%), with high sensitivity for BOB.1 (100%), CD79a (89%) and p63 (82%). CONCLUSIONS: The use of at least three of the most accurate immunohistochemical markers, cyclin E, CD79a and BOB.1, may be helpful in the differential diagnosis of cHL and PMBCL.

Optimized processing of fine-needle lymph node biopsies for automated immunostaining.

A straightforward, reliable technique for postcollection processing and evaluation of cytologic specimens for antigen detection using an automated immunostainer was developed. Visual assessment of cell suspension turbidity was used in parallel with light microscopic examination of concentrated cytospin preparations to verify the diagnostic utility of samples for immunocytochemical staining. Fine-needle lymph node biopsies from 81 dogs with lymphadenomegally and a cytologic or histologic diagnosis of lymphoma were introduced into ethylenediamine tetra-acetic acid tubes containing standardized storage media. Cell suspension turbidity was assessed to estimate cell concentration and resultant volume required for cytospin preparations with optimal cellularity. Preliminary cytospin preparations (using estimated volumes based upon turbidity) were stained using modified Wright stain and examined microscopically for intact neoplastic cell concentration. Once an optimal volume for cytospin preparations was established, additional concentrated slides were prepared for immunophenotyping, using an automated immunostainer and antibodies specific for cluster of differentiation (CD)79a and CD3e. All cell suspension samples with adequate gross turbidity had ample intact neoplastic cell concentration for immunocytochemical staining. Based on CD79a and CD3e expression, 51 (63%) B cell, 19 (23%) T cell, 3 mixed T and B cells (4%), and 3 non-T- and non-B-cell lymphomas (4%), as well as 5 (6%) nondiagnostic samples were identified. Three out of 5 of the nondiagnostic samples were submitted early in the investigation prior to the establishment of gross specimen turbidity guidelines. Immunocytochemical staining results were in complete agreement with all 6 available immunohistochemical correlates. The ability to visually assess sample adequacy prior to sample submission may encourage more widespread use of immunocytochemical techniques.

Can kidney posttransplant lymphoproliferative disorder be detected in voided urine? A case report.

BACKGROUND: Posttransplant lymphoproliferative disorder (PTLPD) is a relatively common complication in kidney transplant recipients. It can involve the kidney allograft itself as well as extragraft sites. It is usually suspected clinically and diagnosed by tissue biopsy. It comprises a spectrum of lesions ranging from lymphoid byperplasia to frank lymphoma. CASE: A 21-year-old male presented with recurrent episodes of microhematuria 2 years after a renal transplant. Voided urine cytology showed anaplastic, discohesive cells. Immunocytochemistry study on a urine cytospin showed the cells to be positive for LCA, CD20 and for CD79a and negative for CD3, CD15, CD30, cytokeratin, S-100 and HMB45, confirming their lymphoid origin. CONCLUSION: This is a rare suspected occurrence of PTLPD in voided urine. The presumptive diagnosis was supported by immunocytochemistry. However, voided urine cytology should not be considered a standard diagnostic test due to its low sensitivity. Diagnosis and proper subclassification of kidney PTLPD should be confirmed by histologic tissue study with supportive ancillary studies--e.g., immunohistochemistry and flow cytometry.

Cutaneous angiogenesis in patient with intravascular lymphoma (IVL): A case report.

We present a case of widespread cutaneous telangiectasias in a patient with a B-cell intravascular lymphoma most likely representing tumor-induced angiogenesis. The patient presented with a rapid onset of large cutaneous telangiectasias and skin edema, followed by the development of multiorgan failure. We describe difficulties with the ante-mortem diagnosis in the patient with predominant, clinically observed, skin lesions. The patient had disseminated disease involving many organs with a rapidly fatal outcome. The final diagnosis of intravascular malignant lymphoma (IVL) was established post-mortem after morphological and immunohistochemical studies of the autopsy material.

A principal case of multiple lymphoid collision tumors involving both B-cell chronic lymphocytic leukemia and metastatic malignant melanoma.

B-cell chronic lymphocytic leukemia/small lymphocytic B-cell lymphoma (CLL/B-SLL) is a neoplasm of B-cell lymphocytes that occurs frequently in the older population as an asymptomatic elevation of the white blood cell count (WBC) and has a good overall prognosis. Malignant melanoma of the skin is a neoplasm derived from cutaneous melanocytes that frequently arises among the elderly and, depending on certain histopathologic features, may metastasize loco-regionally or distally. However, only one report describes synchronous presentation of these two malignancies within the same lymph node. In this report, we present the unique case of an 87-year-old male with a presumed history of indolent CLL/B-SLL, in which metastatic malignant melanoma and CLL/B-SLL both involved 112 of 145 dissected regional lymph nodes. Possible explanations regarding the mechanisms that can lead to this rare presentation of both CLL/B-SLL and melanoma in the same lymph nodes are discussed.

Clinicopathological significance of CD79a expression in classic Hodgkin lymphoma.

Classic Hodgkin lymphoma (CHL) is a lymphoid neoplasia characterized by the presence of large tumor cells, referred to as Hodgkin and Reed-Sternberg (HRS) cells, originating from B-cells in an inflammatory background. As the clinical significance of B-cell markers has yet to be fully elucidated, this study aimed to clarify the clinicopathological significance of CD79a in 55 patients with CHL. They were immunohistochemically divided into two groups, comprising of 20 CD79a-positive and 35 CD79a-negative patients. There was no significant correlation between CD79a and CD20 expression (rs = 0.125, P = 0.362). CD79a-positive patients were significantly older at onset (P = 0.011). There was no significant correlation between CD79a-positivity and clinical stage (P = 0.203), mediastinal involvement (P = 0.399), extranodal involvement (P = 0.749), or laboratory findings, including serum levels of lactate dehydrogenase (P = 1) and soluble interleukin-2 receptor (P = 0.251). There were significant differences in overall survival (OS) (P = 0.005) and progression-free survival (PFS) (P = 0.007) between CD79a-positive and CD79a-negative patients (5-year OS: 64.6% and 90.5%; 5-year PFS: 44.0% and 76.6%, respectively). Five patients in whom the majority (> 80%) of HRS cells expressed CD79a consisted of 4 males and 1 female aged between 52 and 81 years; 4 of them were in a limited clinical stage. We concluded that CD79a-positive CHL may have unique clinicopathological features.

Molecular and phenotypic characterization of an early T-cell precursor acute lymphoblastic lymphoma harboring PICALM-MLLT10 fusion with aberrant expression of B-cell antigens.

T-cell acute lymphoblastic leukemia/lymphoma (T-ALL/LBL) is usually diagnosed based on the presence of immature lymphoid marker terminal deoxynucleotidyl transferase (TdT), and T-cell specific markers, specifically CD3, by immunohistochemistry (IHC) staining on bone marrow and/or extramedullary tissue. We present a novel, TdT and CD3 negative, aggressive early T-cell precursor LBL (ETP-LBL) initially misdiagnosed as a high grade B-cell lymphoma due to expression of CD79a and the erroneous detection of BCL2/IGH fusion. The patient was eventually evaluated using molecular diagnostic techniques, including fluorescence in situ hybridization (FISH) and next generation sequencing (NGS) assays that demonstrated PICALM-MLLT10 fusion and a NOTCH1 mutation in the absence of BCL2/IGH fusion. The use of NGS, specifically mate-pair sequencing (MPseq), subsequently confirmed an in-frame PICALM-MLLT10 fusion. Our retrospective analysis showed that PICALM-MLLT10 fusion has no association with CD3/TdT negativity, as 6/49 T-ALL/LBL cases from Mayo Clinic database (01/1998-09/2018), including this case, were noted to have PICALM-MLLT10 fusion; however, none of the other cases were associated with CD3/TdT negativity. We emphasize the importance of a comprehensive hematopathologic evaluation including multiple molecular studies for the appropriate interrogation and classification of a difficult acute leukemia diagnosis, and to prevent potential diagnostic errors of clinical significance.

Residual inflammation after rituximab treatment is associated with sustained synovial plasma cell infiltration and enhanced B cell repopulation.

OBJECTIVE: To investigate the clinical effects of rituximab treatment in relation to immunological effects of rituximab on tissue-derived B lineage cells and repopulation of circulating B cells. METHODS: A total of 24 patients with rheumatoid arthritis (RA) were treated with 2x1000 mg rituximab and assessed clinically at 4, 12, 18 and 24 weeks using a 44-joint Disease Activity Score (DAS(44)). Synovial biopsies were analysed with immunohistochemistry at baseline and 12 weeks after treatment. Peripheral blood mononuclear cells were analysed by high sensitivity flow cytometry at all timepoints. RESULTS: In this study, a cohort of patients was dichotomised according to those who achieved a low disease activity score (DAS(44)<2.4: LoA group) and those with persistent disease activity (DAS(44)>2.4: HiA group) at any time after rituximab treatment. At baseline, the low activity (LoA) group had significantly lower DAS(44) scores (median 3.33, range 2.84 to 4.23) than the high activity (HiA) group (median 3.73, range 3.03 to 5.23; p = 0.022) and significantly less histological inflammation in synovium (median 6.7, range 1 to 15 vs 16.6, range 4 to 22; p = 0.036). DAS(44) scores before and after rituximab treatment were associated with synovial infiltration of CD79a+ CD20- B cells, morphologically resembling plasma cells. Following treatment with rituximab, the LoA group had significantly reduced repopulation of circulating pre-switched IgD+ B cells (median 0.044%, range 0.002 to 0.66 vs 0.45%, range 0.07 to 9.47; p = 0.006) and post-switched CD27+ B cells (median 0.17%, range 0.04 to 0.39 vs 0.67, range 0.08 to 2.05; p = 0.005) compared to the HiA group. CONCLUSION: The present study demonstrated that a low disease activity state following rituximab was associated with reduced infiltration of CD79a+ CD20- plasma cells in synovium and reduced B cell repopulation.

Comparison of histological characteristics and expression of CD3 and CD79a among the hemal nodes, lymph nodes and spleens of yaks (Bos grunniens).

This experiment compared the histological characteristics and distribution of CD3 and CD79a among yak lymph nodes, hemal nodes and spleen. The three organs from ten clinically healthy newborn and adult yaks were studied using histology and immunohistochemistry. The yak hemal nodes, which consisted of blood sinuses, lymphoid follicles, diffuse lymphoid tissue and lymphoid cords, appeared to share the histological characteristics of the spleen and lymph nodes: the lymphoid follicles of the hemal nodes were much like those of the lymph nodes, which were not surrounded by the central artery and periarteriolar lymphoid sheath. The lymphoid cords of the hemal node, which contained many erythrocytes, were much like the splenic cords. The sinuses of the hemal nodes had a similar structure to the lymph sinuses of the lymph nodes but were engorged with erythrocytes rather than lymph as in the lymph nodes. Interestingly, the splenic sinuses of yak were of two different types: the sinuses with obvious endothelial cells or those consisting of reticular cells. The CD3+ cells were mainly located in the paracortex area and medulla of the lymph nodes, the diffuse lymphoid tissues of the hemal nodes, and the periarteriolar lymphoid sheaths and red pulp of the spleen. Most CD79a+ cells were mainly detected in the lymphoid follicles of all examined lymphoid organs. The results suggested that although the three organs had specific characteristics, in some respects, they had similar organizational structural characteristics and immune functions. These may be useful to better understand the relationship between the morphology and function of these organs and provide useful references for normal yak lymphoid organs.

[Intravascular large B-cell lymphoma: report of two autopsy cases with literature review].

OBJECTIVE: To study the clinicopathologic features of intravascular large B-cell lymphoma (IVLBCL). METHODS: Two autopsy cases of IVLBCL were retrieved from the archival file. The clinicopathologic features, immunohistochemistry and molecular findings were studied. RESULTS: The deceased were 70-year-old and 50-year-old males. Both of them had complained of a sudden onset of weakness and numbness of lower extremities. The clinical course deteriorated rapidly, with multi-organ failure. They died 85 days and 44 days after the presentation, respectively. Post-mortem examination did not reveal any mass lesion, except the presence of multiple skin and epicardium nodules, ranging from 0.5 cm to 2.5 cm in diameter, in the first patient. Pericardial effusion, ascites and pleural effusion were also observed. Histologically, neoplastic lymphoid cells filled up the small vessel lumina in many organs, including brain, hypophysis, spinal cord, spinal nerve roots, heart, lungs, kidneys, liver, spleen, digestive tract, pancreas, adrenal, thyroid, testes and lymph nodes. The tumor cells were relatively monotonous and of medium to large in size with round vesicular nuclei and 1 to 3 small basophilic nucleoli. Immunohistochemical study showed that the lymphoma cells expressed B-cell markers CD20 and CD79a, occasionally positive for CD5 and bcl-2 but negative for CD3, bcl-6, CD10, CD30, myeloperoxidase and cytokeratin. In-situ hybridization for Epstein-Barr virus-encoded RNA was negative. The proliferative index, as demonstrated by Ki-67 staining, was about 80%. Molecular study showed the presence of immunoglobulin heavy chain gene rearrangement in both cases, T-cell receptor-gamma gene rearrangement was not found. CONCLUSIONS: IVLBCL may present as neurological disturbance and carries distinctive morphologic characteristics, immunophenotype and molecular findings. The prognosis of this disease is often dismal.

[Burkitt's lymphoma of the spermatic cord: a case report and review of the literature].

OBJECTIVE: To investigate the clinicopathological characteristics of primary Burkitt's lymphoma (BL) in the spermatic cord. METHODS: A case of BL of the spermatic cord was studied by histopathology and immunohistochemical techniques. The clinical data and the related literature were reviewed. RESULTS: The patient was a 4-year-old boy, who was accidentally found with a bump in the scrotum. Surgery showed it to be a tumor located in the left spermatic cord and 5 cm x 3 cm x 2 cm in size, gray and fish-like on cross-sectional imaging. Histologically, it was characterized by monotonous infiltration of medium-sized cells with round nuclei, coarse chromatin, 2-5 basophilic nucleoli, and an appreciable rim of basophilic cytoplasm, in a typically starry-sky pattern imparted by interspersed tangible-body macrophages. Immunohistochemically, the tumor cells were diffused, positive for CD20 and CD79, some for CD10 and about 95% with the nuclear expression of Ki-67, but negative for CD3, CD43, bcl-2 and TdT as well as for EBER in situ hybridization. CONCLUSION: Primary spermatic cord BL is extremely rare, highly aggressive and with poor prognosis. Diagnosis of the tumor relies on its pathological characteristics and immunohistochemical staining. It is essential to differentiate BL from other types of lymphomas and malignant small-cell tumors of the non-lymphatic system.

Detection of progressive and regressive phase and LINE-1 retrotransposon in transfected dogs with transmissible venereal tumor during chemotherapy.

Canine transmissible venereal tumor (CTVT) is a tumor that commonly occurs in genital and extragenital sites of both genders. Long interspersed nuclear elements (LINE-1) retrotransposon has a pivotal role in allogenic transfection among uncontrolled dog populations. This study aimed to perform pathomorphological, immunohistochemical, and in situ polymerase chain reaction (PCR) evaluation of CTVT (n = 18) in transfected dogs during chemotherapy. Immunohistochemically, tumor phases were investigated by using specific markers (CD3, CD4, CD8, CD79, and transforming growth factor beta [TGF-ß]), and investigated an amplified specific sequence of TVT LINE-1 retrotransposon by in situ PCR. Polyhedral-shaped neoplastic cells that had large, round, hypo/hyperchromatic nuclei and eosinophilic cytoplasm were detected. All marker results were positive, especially in the early weeks of recovery. CD4 and TGF-ß markers were conspicuously positive at the initial stage. In situ PCR LINE-1 sequence was initially positive in only four cases. It is believed that the CD and TGF-ß markers provide phase identification at tumor initiation and during chemotherapy. It is thought that presence of T and B lymphocytes, which have roles in cellular and humoral immunity, is needed so that regression of the tumor is possible.