Presence Of Viable Tumor Cells Research Articles

Histopathological Analysis of Nodal Disease After Chemoradiation Reveals Viable Tumor Cells as the most Important Prognostic Factor in Head and Neck Squamous Cell Carcinoma

BackgroundIn head and neck squamous cell carcinoma (HNSCC), salvage neck dissection (ND) is required after primary chemoradiation in case of residual nodal disease. Upon histopathological examination, viability of tumor cells is assessed but little is known about other prognostic histopathological features. In particular, the presence of swirled keratin debris and its prognostic value is controversial. The aim of this study is to examine histopathological parameters in ND specimens and correlate them with patient outcome to determine the relevant parameters for histopathological reporting.Materials and MethodsSalvage ND specimen from a cohort of n = 75 HNSCC (oropharynx, larynx, hypopharynx) patients with prior (chemo) radiation were evaluated on H&E stains for the following parameters: viable tumor cells, necrosis, swirled keratin debris, foamy histiocytes, bleeding residues, fibrosis, elastosis, pyknotic cells, calcification, cholesterol crystals, multinucleated giant cells, perineural, and vascular invasion. Histological features were correlated with survival outcomes.ResultsOnly the presence / amount (area) of viable tumor cells correlated with a worse clinical outcome (local and regional recurrence-free survival, (LRRFS), distant metastasis-free survival, disease-specific survival, and overall survival, p < 0.05) in both the univariable and multivariable analyses.ConclusionWe could confirm the presence of viable tumor cells as a relevant negative prognostic factor after (chemo) radiation. The amount (area) of viable tumor cells further substratified patients with worse LRRFS. None of the other parameters correlated with a distinctive worse outcome. Importantly, the presence of (swirled) keratin debris alone should not be considered viable tumor cells (ypN0).

Neoadjuvant Cemiplimab for Stage II to IV Cutaneous Squamous-Cell Carcinoma

BackgroundIn a pilot study involving patients with cutaneous squamous-cell carcinoma, a high percentage of patients had a pathological complete response with the use of two doses of neoadjuvant cemiplimab before surgery. Data from a phase 2 study are needed to confirm these findings.MethodsWe conducted a phase 2, confirmatory, multicenter, nonrandomized study to evaluate cemiplimab as neoadjuvant therapy in patients with resectable stage II, III, or IV (M0) cutaneous squamous-cell carcinoma. Patients received cemiplimab, administered at a dose of 350 mg every 3 weeks for up to four doses, before undergoing surgery with curative intent. The primary end point was a pathological complete response (the absence of viable tumor cells in the surgical specimen) on independent review at a central laboratory, with a null hypothesis that a pathological complete response would be observed in 25% of patients. Key secondary end points included a pathological major response (the presence of viable tumor cells that constitute ≤10% of the surgical specimen) on independent review, a pathological complete response and a pathological major response on investigator assessment at a local laboratory, an objective response on imaging, and adverse events.ResultsA total of 79 patients were enrolled and received neoadjuvant cemiplimab. On independent review, a pathological complete response was observed in 40 patients (51%; 95% confidence interval [CI], 39 to 62) and a pathological major response in 10 patients (13%; 95% CI, 6 to 22). These results were consistent with the pathological responses determined on investigator assessment. An objective response on imaging was observed in 54 patients (68%; 95% CI, 57 to 78). Adverse events of any grade that occurred during the study period, regardless of whether they were attributed to the study treatment, were observed in 69 patients (87%). Grade 3 or higher adverse events that occurred during the study period were observed in 14 patients (18%).ConclusionsNeoadjuvant therapy with cemiplimab was associated with a pathological complete response in a high percentage of patients with resectable cutaneous squamous-cell carcinoma. (Funded by Regeneron Pharmaceuticals and Sanofi; ClinicalTrials.gov number, NCT04154943.)

HGG-55. An adolescent with a High-Grade Glioma: persuing a specific diagnosis

BACKGROUND: Recognizing the value of molecular parameters provides evidence with repercussions on diseases classification. CLINICAL CASE: A thirteen-year-old girl was admitted in coma at the Intensive Care Unit due to a hemorrhagic stroke. CT revealed an expansive left frontal lobe lesion with perilesional edema and compression signs by an extensive hematoma. She underwent an emergency decompressive craniotomy for hematoma drainage, and the lesion was biopsied. The subsequent MRI showed a large heterogeneous mass with a cystic component. A first subtotal resection of the lesion was performed considering the language area. The histopathological examination showed tumor cells organized in perivascular pseudorosettes around the central hyalinized thickened blood vessel throughout the tumor, a pattern typically encountered in astroblastoma. Immunochemistry stains revealed a positive reactivity for GFAP, Olig2, and synaptophysin. The molecular study found a BRAFV600E mutation and homozygous CDKN2A e CDKN2B deletions; no TERT amplification, BCORL1, nor MN1 gene rearrangements were identified. She completed six weeks of radiotherapy without neurological signs/symptoms. A new surgical intervention was done 12 weeks later due to a slight increase of residual lesion. The rapid growth and the presence of viable tumor cells justified the therapy with irinotecan and bevacizumab, with mild adverse effects. One year after diagnosis, the adolescent is clinically well, without neurologic deficits and the MRI without evidence of residue or tumor recurrence. DISCUSSION: We began therapy based on morphological diagnosis. Complete resection and radiotherapy proved to be beneficial as in other high-grade gliomas, but the value of other therapies is still unknown. The BRAF gene variant would allow treatment with dabrafenib (in association with trametinib) or vemurafenib; however, she presents molecular findings associated with poor prognosis (CDKN2A deletion) that may impair the effectiveness of these therapies. According to the new WHO classification, the absence of MN1 alterations precludes the diagnosis of astroblastoma.

Abstract No. 131 Evaluation of the Ki-67 labeling index on immediate pre-ablation biopsies as a predictive biomarker of local recurrence of colorectal cancer liver metastases

The presence of residual viable tumor cells in ablation zone biopsies has been shown to predict shorter local tumor progression-free survival (LTPFS). The purpose of this study was to evaluate if the Ki-67 labeling index (LI), as assessed on immediate pre-ablation biopsies of colorectal cancer liver metastases (CLM), is associated with the presence of viable tumor cells in the ablation zone and/or local tumor recurrence.

The effect of direct cell injury inflicted by cryotherapy on eyelid sebaceous gland carcinoma cells: An ex-vivo experimental study.

Purpose:To evaluate the effect of direct cell injury of cryotherapy on eyelid sebaceous gland carcinoma cells by an ex vivo cryotherapy experiment.Methods:It was a prospective interventional case series. Six patients with biopsy-proven nodular sebaceous gland carcinoma were included. After excision of the mass, a thin slice of the mass resembling the thickness of the conjunctiva was shaved off and was oriented over the broad end of a tissue forceps. Cryotherapy was applied to both its anterior and posterior aspects by the triple freeze-thaw technique. The mass was then labeled and sent separately for histopathological evaluation by fixation and staining.Results:A total of six patients with a mean age of 58.2 ± 15.5 years were included. There were four females and two males. The mean duration of the lesion was 21.6 ± 17.51 months. All patients had involvement of the upper eyelid. The patients were clinically staged as T2b (n=2), T1a (n=2), T2c (n=1), and T3a (n=1) respectively. There was no regional lymphadenopathy or metastasis in any of the cases. The experimental cryo-tissue containing the cryo-treated lesion revealed the presence of viable tumor cells (>50%) in all six specimens.Conclusion:The direct cell injury caused by cryotherapy may not be sufficient to kill all the residual sebaceous gland carcinoma cells on the tumor bed.

Occurrence of seeding metastases in resectable perihilar cholangiocarcinoma and the role of low-dose radiotherapy to prevent this.

BACKGROUNDPreoperative biliary drainage in patients with presumed resectable perihilar cholangiocarcinoma (PHC) is hypothesized to promote the occurrence of seeding metastases. Seeding metastases can occur at the surgical scars or at the site of postoperative drains, and in case of percutaneous biliary drainage, at the catheter port-site. To prevent seeding metastases after resection, we routinely treated PHC patients with preoperative radiotherapy (RT) for over 25 years until January 2018.AIMTo investigate the incidence of seeding metastases following resection of PHC.METHODSAll patients who underwent resection for pathology proven PHC between January 2000 and March 2019 were included in this retrospective study. Between 2000-January 2018, patients received preoperative RT (3 × 3.5 Gray). RT was omitted in patients treated after January 2018.RESULTSA total of 171 patients underwent resection for PHC between January 2000 and March 2019. Of 171 patients undergoing resection, 111 patients (65%) were treated with preoperative RT. Intraoperative bile cytology showed no difference in the presence of viable tumor cells in bile of patients undergoing preoperative RT or not. Overall, two patients (1.2%) with seeding metastases were identified, both in the laparotomy scar and both after preoperative RT (one patient with endoscopic and the other with percutaneous and endoscopic biliary drainage).CONCLUSIONThe incidence of seeding metastases in patients with resected PHC in our series was low (1.2%). This low incidence and the inability of providing evidence that preoperative low-dose RT prevents seeding metastases, has led us to discontinue preoperative RT in patients with resectable PHC in our center.

Immediate post-thermal ablation biopsy of colorectal liver metastases to predict oncologic outcomes.

4602 Background: Thermal ablation (TA) is used as a local cure for selected colorectal liver metastases (CLM) with minimal risk. A critical limitation of TA has been early local tumor progression (LTP). The goal of this study is to establish the role of ablation zone (AZ) biopsy in predicting LTP. Methods: This institutional review board-approved prospective study included patients with CLM of 5cm or less in maximum diameter, with confined liver disease or stable, limited extrahepatic disease. Both radiofrequency(RF) and microwave(MW) ablation modalities were used. A biopsy of the center and margin of the AZ was performed immediately after ablation. The applicators were also examined for the presence of viable tumor cells. All samples containing morphologically identified tumor cells were further interrogated with immunohistochemistry to determine the proliferative and viability potential of the detected tumor cells. Ablation margin size was evaluated on the first CT scan performed 4–8 weeks after ablation and was confirmed by 3D assessment with Ablation Confirmation Software (Neuwave™). Variables were evaluated as predictors of time to LTP with the competing-risks model (uni- and multivariate analyses). Results: Between November 2009 and February 2019, 102 patients with 182 CLMs were enrolled. Mean tumor size was 2.0 cm (range, 0.6–4.8 cm). MW was used in 95/182 (52%) tumors and RF in 87/182 (48%). Median follow-up was 19 months. Technical effectiveness was evident in 178/182 (97%) ablated tumors on the first contrast material–enhanced CT at 4–8-weeks post-ablation. The cumulative incidence of LTP at 12 months was 19% (95% confidence interval [CI]: 14, 27). Samples from 64 (35%) of the 178 technically successful cases contained viable tumor. At univariate analysis, tumor size, minimal margin size, and biopsy results were significant in predicting LTP. In a multivariate model, margin size of less than 5 mm (P &lt; .001; hazard ratio [HR], 4.3), and positive biopsy results (P = .02; HR, 1.8) remained significant. LTP within 12 months after TA was noted in 3% (95% CI: 1, 6) of tumor-negative biopsy CLMs with margins of at least 5 mm. Conclusions: Biopsy and pathologic examination of the AZ predicts LTP regardless of TA modality used. This can optimize ablation as a potential local cure for patients with limited CLM.

Correlation between CT morphologic appearance and histologic findings in colorectal liver metastasis after preoperative chemotherapy.

Radiological evaluation of the efficacy of preoperative chemotherapy for colorectal liver metastasis (CRLM) is the most important tool for determining treatment strategies. The aim of this study was to identify a correlation between morphologic appearance on computed tomography (CT) and histologic findings of CRLM after preoperative chemotherapy. We examined 47 patients who had undergone a first hepatic resection for CRLM after preoperative chemotherapy and had received contrast-enhanced CT scans. We assessed the morphologic appearance of the overall attenuation based on metastases changing from heterogeneous to mixed and homogenous lesions, the tumor-liver interface, and the peripheral rim enhancement on CT. Histologic parameters included usual necrosis (UN), infarct-like necrosis (ILN), three-zonal change, dangerous halo, mucous lake, shape of ILN, dominant type of necrosis, and presence of viable tumor cells. The relationship between morphologic appearance and histologic findings was evaluated. CT overall attenuation revealed that UN predominance was more common in the heterogeneous group than in the mixed and homogeneous groups (P = 0.011). The frequency of ILN increased sequentially from ill-defined to variable and sharp at the tumor-liver interface (P = 0.038), and the frequency of UN decreased sequentially from present to partially resolved and completely resolved in the peripheral rim enhancement (P = 0.023). The histologic presence of viable tumor cells was closely associated with the tumor-liver interface (P = 0.0003) and the peripheral rim enhancement (P = 0.004). CT morphologic appearance of CRLM after preoperative chemotherapy is correlated with histologic findings regarding necrosis.

Colorectal Cancer Liver Metastases: Biopsy of the Ablation Zone and Margins Can Be Used to Predict Oncologic Outcome.

Purpose To establish the prognostic value of biopsy of the central and marginal ablation zones for time to local tumor progression (LTP) after radiofrequency (RF) ablation of colorectal cancer liver metastasis (CLM). Materials and Methods A total of 47 patients with 67 CLMs were enrolled in this prospective institutional review board-approved and HIPAA-compliant study between November 2009 and August 2012. Mean tumor size was 2.1 cm (range, 0.6-4.3 cm). Biopsy of the center and margin of the ablation zone was performed immediately after RF ablation (mean number of biopsy samples per ablation zone, 1.9) and was evaluated for the presence of viable tumor cells. Samples containing tumor cells at morphologic evaluation were further interrogated with immunohistochemistry and were classified as either positive, viable tumor (V) or negative, necrotic (N). Minimal ablation margin size was evaluated in the first postablation CT study performed 4-8 weeks after ablation. Variables were evaluated as predictors of time to LTP with the competing-risks model (uni- and multivariate analyses). Results Technical effectiveness was evident in 66 of 67 (98%) ablated lesions on the first contrast material-enhanced CT images at 4-8-week follow-up. The cumulative incidence of LTP at 12-month follow-up was 22% (95% confidence interval [CI]: 12, 32). Samples from 16 (24%) of 67 ablation zones were classified as viable tumor. At univariate analysis, tumor size, minimal margin size, and biopsy results were significant in predicting LTP. When these variables were subsequently entered in a multivariate model, margin size of less than 5 mm (P < .001; hazard ratio [HR], 6.7) and positive biopsy results (P = .008; HR, 3.4) were significant. LTP within 12 months after RF ablation was noted in 3% (95% CI: 0, 9) of necrotic CLMs with margins of at least 5 mm. Conclusion Biopsy proof of complete tumor ablation and minimal ablation margins of at least 5 mm are independent predictors of LTP and yield the best oncologic outcomes. (©) RSNA, 2016.

Abstract 735: Evaluation of cabozantinib in combination with abiraterone, enzalutamide or docetaxel in a castration-resistant prostate cancer xenograft in vivo

Abstract New second-generation hormonal therapy agents abiraterone (abi) and enzalutamide (enz), and standard chemotherapy docetaxel (doc) provide survival benefits for patients with castration-resistant prostate cancer (CRPC). However, despite these survival benefits, CRPC is still an incurable disease. Therefore, combination treatments with other agents are under consideration to potentially improve patient outcome. Cabozantinib (cabo), an inhibitor of tyrosine kinases including MET, VEGFR2, RET, KIT and AXL, has shown activity as monotherapy in clinical investigations in CRPC. Our objective was to evaluate effects of cabo in combination with abi, enz or doc on a CRPC xenograft tumors in vivo. Mice with subcutaneous LuCaP 35CR CRPC patient-derived xenograft tumors were treated with abi (0.5 mmol/kg), enz (50 mg/kg), doc (10 mg/kg) or cabo (10 mg/kg) monotherapies or with combinations of cabo with abi, enz or doc. Mice were treated for 4 weeks (wks) and followed for additional 6 wks. After 4 wks of treatment abi and enz exhibited limited activity in this model (∼20% inhibition of tumor volume (TV), not significant), doc exhibited ∼40% TV inhibition (P=0.017), and cabo exhibited ∼60% TV inhibition (P&amp;lt;0.001) vs untreated tumors. Combination of cabo with each of these agents provided reduction in TV vs single agent treatments: ∼40% inhibition vs abi only, P=0.0039; ∼60% inhibition vs enz only, P&amp;lt;0.0001; and ∼70% inhibition vs doc only, P&amp;lt;0.0001. When the tumors were followed for 6 wks after discontinuation of the treatment, all treated tumors started to grow at rates similar to the untreated tumors, indicating presence of viable tumor cells. At the end of the study (10 wks) there were survival benefits for monotherapies, and further survival benefits were detected for combination of enz+cabo vs enz as a monotherapy (medium survival 5.7 vs 3.5 wks, P=0.0087) and doc+cabo vs doc as monotherapy (medium survival 4.75 vs 3.5 wks, P=0.0003), however survival of animals treated with the combinations was not significantly different from the survival of animals treated with cabo monotherapy. In the second study, where mice were treated for up to 18 wks, there were significant survival benefits detected for enz+cabo vs enz (median survival 15.5 vs 6.5 wks; P=0.0039), and for doc+cabo vs doc (median survival 15 vs 6.8 wks; P=0.0008); however, the cabo combinations with enz and doc had survival times similar to cabo monotherapy (16 wks). No significant effects on survival were detected for abi+cabo vs abi as monotherapy. Cabo at 10 mg/kg administered long term did not result in decreased body weight in comparison to non-treated mice. These preliminary data indicate that combinations of cabo with enz, doc or abi are active in the subcutaneous LuCaP 35CR xenograft model. Further analyses of the data and tumors are ongoing. Citation Format: Holly M. Nguyen, Lisha G. Brown, Jessica L. Olson, Dana T. Aftab, Robert L. Vessella, Eva Corey. Evaluation of cabozantinib in combination with abiraterone, enzalutamide or docetaxel in a castration-resistant prostate cancer xenograft in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 735. doi:10.1158/1538-7445.AM2014-735

Flow cytometry assessment of residual melanoma cells in tumor‐infiltrating lymphocyte cultures

Adoptive transfer of tumor-infiltrating lymphocytes (TIL) is in development for the treatment of metastatic melanoma. In phase II clinical trials, patients with metastatic melanoma that received TIL after preconditioning had a 50-70% clinical response rate. The current approach to generate TIL is to culture melanoma enzyme digests in the presence of IL-2 for a 10- to 20-day period followed by 2 weeks of rapid expansion (REP). Prior to administration, cell therapies are characterized and tested for purity. TIL are characterized by CD3 surface marker expression, and purity is assessed by the amount of tumor remaining in culture. Evaluating TIL purity has traditionally been done by immunohistochemistry, which is often considered semiquantitative. To generate a quantitative assay, we used multiparameter flow cytometry to evaluate the presence of viable tumor cells by staining TIL populations with a viability dye and an antibody cocktail that detects intracellular tumor-antigens gp100, Mart-1, tyrosinase, S100, and surface tumor-antigen melanoma chondroitin sulfate proteoglycan (MCSP), and CD3 on T cells. Tumors were identified by gating on the viable CD3(-) population. Antigens in tumors were initially optimized with individual antibodies using both immunohistochemistry and flow cytometry. When eight different tumor cell lines were spiked into an activated T cell culture, flow cytometry was able to distinguish lymphocytes from tumors in all samples tested. Most importantly, the assay was able to detect melanoma cells in all enzyme digests (9/9) from patient samples. After IL-2-induced TIL expansion, there was a significant decrease in tumor cells; tumor cells were detected in only 2 of 12 samples. In eight IL-2-induced TIL samples that were further expanded in REP, no tumor cells were detected. We have demonstrated that flow cytometry is an alternative to immunohistochemistry for defining the purity of a TIL population.

Cisplatin-based chemotherapy followed by surgery for malignant nonseminomatous germ cell tumor of mediastinum: one institution’s experience

The objective of this study was to evaluate the efficacy and safety of cisplatin-based chemotherapy followed by surgery for patients with a malignant nonseminomatous germ cell tumor (NSGCT) of the mediastinum. Ten patients with malignant NSGCTs received cisplatin-based induction chemotherapy and then underwent surgery. The clinicopathological characteristics of these 10 patients were examined retrospectively. A partial response to induction chemotherapy was noted in eight patients and no response in two. The induction chemotherapy was tolerated well by all the patients. Each patient underwent complete surgical resection of the residual tumor following chemotherapy. A yolk sac tumor was detected in one patient and malignant teratoma along with a yolk sac tumor in one patient postoperatively. The overall survival of the 10 patients was 67% at 60 months of follow-up. The survival rate at 60 months was poorer for the patients whose resected specimens exhibited the presence of viable cells than for those whose specimens were free of viable cells. Postchemotherapy surgical resection of the residual tumor plays an integral role in the management of patients with NSGCT. The presence of viable tumor cells in the resected specimens is associated with poor survival.

Complete radiological response of colorectal liver metastases (LM) after chemotherapy: Does it mean cure?

3501 Background: Most of patients with colorectal LM receive systemic chemotherapy. Complete response on imaging is considered a good indicator of the efficacy of chemotherapy. Whether complete radiological response is correlated to complete pathological response is not well known. The objective of this study was to determine the significance of complete radiological response of LM after chemotherapy. Methods: Between 1998 and 2004, 586 patients were treated in one institution for colorectal LM. 38 with the following criteria were included in the study: less than 10 LM before chemotherapy, disappearance of one or several LM on helical CT scan and ultrasound after chemotherapy, surgery with liver examination and intraoperative ultrasound within 4 weeks after imaging, no extra-hepatic disease, follow-up at least one year after surgery. Results: Overall, 66 LM disappeared on imaging after chemotherapy (38 patients). In 9 patients, persistent macroscopic disease was observed at surgery at the site of 20/66 (30%) LM although there were considered in complete response on imaging. In 15 patients, with no remnant disease at surgery, the sites of 15 initial LM were resected. Pathological examination of theses sites of LM considered in complete response showed the presence of viable tumor cells in 12/15 cases (80%). In 14 other patients, the sites of 31 initial LM, which were not visible at surgical exploration were left in place during surgery. After one year of follow-up, 23/31 (74%) LM considered in complete response had recurred in situ (11/14 patients). Overall, persistent macroscopic or microscopic residual disease or early recurrence in situ were observed in 55/66 LM (83%) with complete radiological response (32/38 patients). Conclusions: Complete radiological response in patients who have received chemotherapy is a useful tool to evaluate the effect of chemotherapy but it does not mean cure in most patients. This should be taken into account when considering multimodality treatment including chemotherapy and surgery. No significant financial relationships to disclose.

Dynamic contrast-enhanced magnetic resonance imaging in radiotherapeutic efficacy in the head and neck tumors

Purpose The purpose to this study is to identify correlations between pathology and dynamic contrast-enhanced magnetic resonance imaging (MRI) and to assess the utility of this technique in the evaluation of radiation response for head and neck cancer. Materials and methods MRI was prospectively performed after radiotherapy in 27 patients with various head and neck tumors. After bolus injection of contrast material, a dynamic study was performed using a spoiled gradient–recalled imaging sequence. The maximum slope of increase (MSI) on the time-intensity curve was displayed as a color-coded image. The ratio of MSI (MSIR) was obtained for tumor and normal muscles. Pathological specimens were obtained after MRI in all cases. Histological grading of irradiation changes was classified into 5 grades (0–4). Correlations between MSIR and histological grade were examined. Results Histologically, 18 tumors were classified as grade 2 (presence of viable tumor cells), 4 were grade 3 (nonviable tumor cells), and 5 were grade 4 (no tumor cells). Although the mean ± SD of MSIR in patients with histological grade 2 or 3 was 7.4 ± 7.9, MSIR in patients with grade 4 was 1.8 ± 0.73, representing a significant difference ( P < .05). Every patient with grade 4 displayed an MSIR of 2.5 or less, although 5 of 22 patients with grade 2 or 3 had an MSIR of 2.5 or less. Conclusions MSI quantitatively reflects response to radiotherapy for head and neck cancer. Color-coded MSI display is feasible for depicting permeability changes after radiotherapy.

In vivo photodynamic effects of phthalocyanines in a skin-fold observation chamber model: role of central metal ion and degree of sulfonation.

Six sulfonated metallophthalocyanines, chelated with either aluminum or zinc and sulfonated to different degrees, were studied in vivo for their photodynamic activity in a rat skin-fold chamber model. The chamber, located on the back of female WAG/Rij rats, contained a syngeneic mammary carcinoma implanted into a layer of subcutaneous tissue. Twenty-four hours after intravenous administration of 2.5 mumol/kg of one of the dyes, the chambers received a treatment light dose of 600 J/cm2 with monochromatic light of 675 nm at a power density of 100 mW/cm2. During light delivery and up to a period of 7 days after treatment, vascular effects of tumor and normal tissue were scored. Tumor cell viability was determined by histology and by reimplantation of the chamber contents into the skin of the same animal, either 2 h after treatment or after the 7 day observation period. Vascular effects of both tumor and subcutaneous tissue were strongest with dyes with the lowest degree of sulfonation and decreased with increasing degree of sulfonation. Tumor regrowth did not occur with aluminum phthalocyanine mono- and disulfonate and with zinc phthalocyanine monosulfonate. With the protocol that was used, complete necrosis without recovery was only observed when reimplantation took place at the end of the 7 day follow-up period. Reimplantation 2 h after treatment always resulted in tumor regrowth. At this interval, the presence of viable tumor cells was confirmed histologically. In general tumor tissue vasculature was more susceptible to photodynamic damage than vasculature of the normal tissue. The effect on the circulation of both tumor and normal tissue increased with decreasing degree of sulfonation.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum squamous cell carcinoma antigen in the monitoring of radiotherapy treatment response in carcinoma of the cervix

In this study, squamous cell carcinoma antigen (SCC) was detected in 96 of 157 patients with squamous cell carcinoma of the cervix and the percentage of patients with raised SCC levels increased with the stage of disease ( P < 0.01). The use of serial SCC assays and cervical biopsy histology during the course of radiotherapy to predict tumor response to irradiation was assessed. In patients who were given external irradiation before intracavitary radium, a high SCC level or the presence of viable tumor cells in the biopsy was found to be of no predictive value. However, at completion of radiotherapy, i.e., after intracavitary radium application, patients with persistently high SCC levels had a significantly higher incidence of residual tumor than patients whose SCC levels returned to normal ( P < 0.01). In 60% of patients with a persistently high SCC level, viable tumor was found in the cervical biopsy at the end of radiotherapy. On the other hand, only 5.4% of patients whose SCC level returned to normal had residual tumor.

Management of choroidal malignant melanoma at Iowa.

Approximately 50% of patients undergoing enucleation for choroidal melanoma will eventually die of metastases. The majority of tumor related deaths will occur within 5 years of enucleation, but late metastases may occur 20-30 years after enucleation. Long-term follow-up of patients refusing treatment has shown a 100% mortality. Although recent reports have attempted to link the performance of enucleation with the development of metastases, examination of available data suggests that metastases occur preoperatively, but are subclinical at the time of enucleation. The most important variable determining prognosis in these cases is the size of the tumor at the time of diagnosis and treatment. Patients who undergo enucleation before the tumor exceeds 7 X 7 X 2 mm have an extremely favorable prognosis. Patients whose maximum tumor diameter exceeds 12 mm have an extremely poor prognosis. Metastases and extrascleral extension have both been reported to occur in tumors which failed to demonstrate growth while being followed. For these reasons we recommend immediate enucleation be carried out in all patients whose tumors exceed 7 X 7 X 2 mm and in whom the diagnosis can be confirmed by indirect ophthalmoscopy, ultrasound, fluorescein angiography, and other tests. Follow-up of non-enucleation methods of treatment of choroidal melanomas is too short to draw meaningful conclusions. The consistent finding of persistence of some tumor mass and the histopathologic presence of viable tumor cells following these treatment modalities leaves us very concerned about the long-term results in these patients. Late metastases continue to occur decades following diagnosis of a choroidal melanoma, especially if it is small. The ophthalmologist must remain aware that the results of his decision concerning treatment of a melanoma may not become apparent for many years.

Transient lysozymuria during Sarcoma I tumor rejection

The present studies on the in vivo regulation of lysozyme synthesis by cytotoxic macrophages infiltrating Sarcoma I (Sa I) tumor ascites of C57BL/KS mice are an extension of previous observations that the elevation of serum and urinary lysozyme levels might reflect the degree of macrophage-mediated host resistance. Cytotoxic macrophages of host origin are required in the natural rejection of tumors by C57BL/Ks mice challenged with Sa I. The amounts of lysozyme in the serum and urine of the affected mice are quantitated during Sa I growth and rejection. The enzyme levels are correlated with the number of macrophages in the tumor and with the capacity for lysozyme synthesis measured in vitro on monolayers of macrophages isolated from the ascites. The results indicate that the elevation of serum and urinary lysozyme activity is a reflection of both the increased number of cytotoxic macrophages in the host and the qualitative change in the functional state of the cells. These parameters alone cannot account for the extent of the in vivo release of lysozyme in the serum and urine of the affected animals. High levels of lysozyme are present in the extracellular fluids of the mice only when the macrophages are actively participating in host defense. We conclude that in vivo control mechanisms which depend on the presence of viable tumor cells are responsible for the regulation of lysozyme release by cytotoxic macrophages. However, the high levels of lysozyme do not always indicate complete rejection, as indicated by a preliminary observation in which extensive lysozymuria was followed by tumor relapse.

Cell-mediated immune response to syngeneic ultraviolet-induced tumors: II. The properties and antigenic specificities of cytotoxic T lymphocytes generated in vitro following removal from syngeneic tumor-immunized mice

The immune responses of C3Hf mice to syngeneic fibrosarcomas induced with either ultraviolet light or methychlolanthrene (MCA) were measured in vitro by the ability of cytotoxic lymphocytes (CTL) from immunized animals to kill 51Cr-labeled tumor targets in a 6-hr assay. The CTL were generated by the in vitro culturing of draining popliteal lymph node (DLN) cells derived from animals that were footpad immunized 8 days previously. It was determined that CTL activity could be generated using DLN from both normal (uv tumorresistant) and uv-exposed (uv tumor-susceptible) C3H mice. The kinetics of CTL generation between these two groups, however, was different in that the lymphocytes from normal animals appeared to differentiate into CTL faster than the lymphocytes from the uv-irradiated mice. The in vitro generation of CTL activity was found to be extremely radiosensitive and was also inhibited by the presence of viable tumor cells within the cell culture. Once generated, it was observed that the CTL were extremely insensitive to the effects of gamma irradiation. It was also established that the CTL is a T lymphocyte that appears to be Ia −. The CTL derived from mice immunized to syngeneic uv- or MCA-induced tumors were capable of expressing cross-reactive non-MHC-restricted killing of multiple tumor targets. Cold cell inhibition experiments confirmed the presence of cross-reactive determinants on various tumors and also established the presence within a single CTL preparation of effector cells with specificity for both the unique tumor specific transplantation antigens as well as the common (cross-reactive) tumor-associated antigens.

Dissociation between cytotoxic activity and delayed hypersensitivity against syngeneic or allogeneic tumor grafts.

Cytotoxicity against tumor-specific antigen and allogeneic antigens in mice was raised by immunization with viable tumor cells in saline, but not by immunization with tumor cells in complete or incomplete Freund's adjuvant. On the other hand, strong delayed hypersensitivity as demonstrated by macrophage migration inhibition was detected in the mice immunized with tumor cells in complete or incomplete Freund's adjuvant in both syngeneic and allogeneic systems. After immunization with viable tumor cells, delayed hypersensitivity was raised to a moderate degree in allogeneic, but not at all in syngeneic mice. Cytotoxicity was retained after a booster with syngeneic tumor cells in complete Freund's adjuvant in the mice which had been immunized with viable tumor cells. Cytotoxicity was raised after a booster with viable syngeneic tumor cells in the mice which had been immunized with tumor cells in complete or incomplete Freund's adjuvant. Thus, the development of cytotoxicity depended entirely on the presence of viable tumor cells as an immunogen. Activation of helper T cells was not found in the syngeneic mice immunized with viable tumor cells or tumor cells in complete or incomplete Freund's adjuvant. In allogeneic mice, the activity was facilitated slightly by immunization with viable tumor cells or tumor cells in complete Fruend's adjuvant but not with tumor cells in incomplete Freund's adjuvant. This sytem may be useful for analytical studies on cellular immunity, since cytotoxicity, delayed hypersensitivity, and helper cell activation can be induced in dissociated forms.