Radioimmunoguided surgery of colorectal carcinoma with an 111In-labelled anti-TAG72 monoclonal antibody

Radioimmunoscintigraphy (RIS) and radioimmunoguided surgery (RIGS) were assessed for their usefulness in patients with colorectal carcinoma. Twenty-nine patients (18 primary tumours, 10 with a suspicion of recurrence and one colonic diverticulitis) were studied. Radioimmunoscintigraphy was performed 48 and 72 h after the injection of an anti-TAG72 monoclonal antibody (CYT-103) labelled with 111In. Radioimmunoguided surgery was performed between 72 and 96 h post-injection. During surgery, a systematic screening was performed with a hand-held gamma detecting probe and a surgical index (tumour-to-normal tissue) was obtained. There were statistically significant differences between counts in normal tissue versus tumour (P < 0.001) and RIGS was considered positive for the detection of tumour if the ratio between the counts in the area suspicious of tumour and the counts in the normal tissue was greater than 1.5. The overall sensitivity for RIS and RIGS was 71.4% (55.6% in primary tumours and 100% in recurrences) and 82.1% (83.3% in primary tumours and 80% in recurrences), respectively. Radioimmunoguided surgery changed the surgical procedure in two cases with small tumour deposits. Occult regional lymph node involvement in primary tumours was not found; therefore, RIGS, as a complementary technique to RIS, is particularly useful in recurrences and can help the surgeon in the resection of small tumour deposits which are difficult to localize.     

Imaging of medullary carcinoma of the thyroid using 111In-labelled anti-CEA monoclonal antibody fragments.

The distribution of 111In-labelled anti-carcinoembryonic antigen (CEA) monoclonal antibody fragments [F(Ab')2] was studied in five patients either with known inoperable medullary carcinoma of the thyroid (MCT) or evidence of recurrence/metastases based on elevated calcitonin (hCT) levels. All five cases had elevated serum CEA levels and positive immunohistochemical stains for both hCT and CEA prior to scintigraphy. In two patients with identified inoperable disease both planar and SPECT scans were positive. In the remaining three patients, where the recurrence/metastatic sites were unknown, SPECT images were positive in two. Of these, only one had positive planar images. These results indicate that 111In-labelled anti-CEA F(ab')2 scintigraphy, especially in conjunction with SPECT, is useful for the diagnostic evaluation of patients with MCT. The limiting factor of this technique is the high level of non-specific uptake, particularly in the liver, but improvements in the specificity of newer anti-CEA antibodies and the ability to label these with 99Tcm is addressing this problem.     

Targeting of lymphocytes with 111In-labelled monoclonal antibodies

In this paper, we emphasize the rationale and work-up studies for using two radiolabelled anti-lymphocyte monoclonal antibodies for in vivo application as radiolabelling agents for T and B cells. In vitro experimental work involved radioimmunoassays on human lymphoid cell lines and anticoagulated whole blood with identification of relevant binding kinetics in terms of antibody internalization and elution. We tested also for the effect of the radiolabelled monoclonal antibodies on in vitro cell function defined as mitogen-induced proliferation in whole blood. As a final work-up in an animal model, the distribution of both unlabelled and 111In-labelled anti-Pan T cell monoclonal antibody was studied in the rat. Results from the in vitro experiments pointed to the possibility of using the described technique for specific lymphocyte radiolabelling. The in vivo application enabled us to identify optimal doses of antibody and radioactivity which showed agreement with the in vitro data.     

Imaging of primary and metastatic colorectal cancer using an 111In-labelled antitumour monoclonal antibody (791T/36)

The monoclonal antitumour antibody 791T/36, previously shown to localize in colorectal cancer when radiolabelled with 131I, has been successfully labelled with 111In. A preliminary evaluation of its imaging potential has been made in fourteen patients with primary and secondary colorectal cancer. Positive localization was achieved in tumour sites of all but one of the patients. The sites of uptake of the 111In-labelled antibody were more clearly defined than when using the same antibody labelled with 131I. The mean tumour to normal tissue uptake ratio per gram of tissue from four resected specimens was 2.8:1. Labelling of the antibody did not affect the tumour uptake of the antibody but facilitated interpretation directly from images without the need for background subtraction.     

The preparation and characterisation of 111In-labelled 791T/36 monoclonal antibody for tumour immunoscintigraphy

The anti-human tumour monoclonal antibody 791T/36 was conjugated to the cyclic dianhydride of DTPA and radio-labelled with 111In. The labelling method proved to be both simple and reliable and would be suitable for routine clinical use. Subsequent characterisation of this radio-pharmaceutical in vitro and in tumour-bearing hosts gave a strong indication as to its suitability for clinical tumour localisation studies.     

The preparation and characterisation of 111In-labelled 791T/36 monoclonal antibody for tumour immunoscintigraphy

The anti-human tumour monoclonal antibody 791T/36 was conjugated to the cyclic dianhydride of DTPA and radio-labelled with ¹¹¹In. The labelling method proved to be both simple and reliable and would be suitable for routine clinical use. Subsequent characterisation of this radio-pharmaceutical in vitro and in tumour-bearing hosts gave a strong indication as to its suitability for clinical tumour localisation studies.     

Radionuclide imaging of primary renal-cell carcinoma by I-131-labeled antitumor antibody

A goat antibody against human renal-cell carcinoma reacted on immunofluorescence with renal-cell carcinomas from 20 patients, but not with normal adult human tissues, including kidney. After i.v. administration the I-131-linked antibody showed preferential tumor localization in six of seven patients with primary renal carcinoma. Labeled antitumor antibodies may have the specificity for tumor imaging that current radiopharmaceuticals lack.     

131I标记抗NRP-1单克隆抗体A6在荷瘤裸鼠体内分布和显像研究

目的制备131I标记的抗神经纤毛蛋白-1（NRP-1）单克隆抗体A6（131I-A6）,探讨其作为NRP-1靶向显像新型分子探针的可行性。方法（1）采用Iodogen法对A6进行131I标记,检测其标记率、放化纯和体外稳定性。（2）以人胶质瘤细胞株U87MG为实验细胞进行体外实验,测定131I—A6生物学活性、结合率及其与受体的亲和力。（3）将荷U87MG胶质瘤模型裸鼠采用随机抽样法分为5组,每组5只,分别于注射1．2MBq 131I-A6后24、48、72、96和120h处死,计算各脏器放射性摄取（％ID／g）、肿瘤／血液（T／B）和肿瘤／肌肉（T／M）比值。（4）取6只荷瘤裸鼠,以随机抽样法分为未阻断组和竞争阻断组,前组注射3．7MBq 131I-A6;后组注射3．7MBq 131I-A6和未标记的700仙gA6,均分别于注射后24、48、72、96和120h行SPECT／CT显像。采用两样本t检验对实验数据进行统计学分析。结果（1）131I—A6标记率为（95．46±3．34）％,放化纯〉95％;131I—A6在室温下PBS溶液中放置至96h,其放化纯仍〉85％。（2） 131I-A6与U87MG胶质瘤细胞特异性结合率1h达到最高值,为（15．80±1．30）％,在加入未标记的抗体A6时,U87MG细胞对131I-A6明显受抑制（t=2．862,P〈0．05）;与细胞表面抗原的亲和力（kd）为（1．67±0．14）nmol／L。（3）24h时131I—A6在荷瘤裸鼠血液放射性最高,为（8．00±1．42）％ID／g;其次是肝脏和肿瘤组织,分别为（7．68±1．56）和（6．00±1．24）％ID／g;脑、骨、肌肉组织放射性计数较低。给药后24h,T／B和T／M分别为0．78±0．10和3．20±0．30,随时间延长比值逐渐增高,在120h达到最高,分别为1．87±0．50和7．13±0．24。（4）体内显像示,注射 131I-A6后24h肿瘤略显影,随时间延长变清晰,120h显影为最清晰,阻断后未见肿瘤显影。结论 131I-A6的标记方法简单易行,标记率高,产物稳定性好,?     

Diagnosis of deep venous thrombosis of the leg using immunoscintigraphy with 111In-labelled monoclonal antifibrin antibody fragments

Fourteen unselected consecutive patients with clinically suggested deep venous thrombosis were studied using 111In-labelled monoclonal antibody Fab-fragments (59D8) in order to evaluate the accuracy of antifibrin imaging. Scintigrams of the legs were obtained at 1 to 4 h and 20 to 28 h in all patients and in seven dynamic studies were also performed. Contrast medium phlebography used as diagnostic standard revealed thrombosis in six patients who also had increased uptake of the tracer at immunoscintigraphy. Three patients had a positive finding at immunoscintigraphy without thrombosis. Because of the high sensitivity of this method, normal antifibrin images may be used to exclude thrombosis. Phlebography may then be restricted to positive cases to confirm the presence of thrombosis.     

A comparative distribution study of 111In-labelled DTPA and TTHA monoclonal antibody conjugates in a choriocarcinoma xenograft model

Conjugates of the chelating agents DTPA and TTHA with a monoclonal anti-HCG were prepared. The tissue distribution of the ¹¹¹In-labelled conjugates and also ¹¹¹In-citrate was studied in mice bearing human choriocarcinoma xenografts. The antibody conjugates both gave high liver and spleen radionuclide accumulation. Elevated femur levels were observed for the TTHA conjugate and ¹¹¹In-citrate. Generally the DTPA conjugate showed the highest tumor/tissue ratios, although its tumor/blood ratio was lower than the other two materials. The results infer that the DTPA conjugate has the greatest utility as an imaging agent but that it would require a background subtraction technique.     

Clinical experience with intra lymphatic administration of111In-labelled monoclonal antibody PAY 276 for the detection of pelvic nodal metastases in prostatic carcinoma

The ability of¹¹¹In-PAY 276 (anti prostatic acid phosphatase antibody) in detecting pelvic lymph node metastasis following bipedal intra lymphatic administration was studied in five patients with carcinoma of the prostate. The labeled antibody was injected directly into the lymphatics of each foot. Planar and tomographic images radioactivity content of lymph nodes resected during staging pelvic lymphadenectomy were compared to the histologic and immunoperoxidase findings. Radioactivity in pelvic lymph nodes was prominently seen within 20 min of injection and was present 16 days later. Persistent accumulation of tracer in the lymphatics of the lower extremities was also observed in all patients 16 days post injection. Radioactivity counts in tumor-free lymph nodes were higher than in tumored lymph nodes resected. Our results demonstrate that intra lymphatic administration of¹¹¹In-labeled PAY 276 monoclonal antibody has major technical limitations, and that further research directed at the causes of tracer accumulation in the lymphatics and tumor-free lymph nodes is required.     

Clinical experience with intra lymphatic administration of 111In-labelled monoclonal antibody PAY 276 for the detection of pelvic nodal metastases in prostatic carcinoma

The ability of 111In-PAY 276 (anti prostatic acid phosphatase antibody) in detecting pelvic lymph node metastasis following bipedal intra lymphatic administration was studied in five patients with carcinoma of the prostate. The labeled antibody was injected directly into the lymphatics of each foot. Planar and tomographic images radioactivity content of lymph nodes resected during staging pelvic lymphadenectomy were compared to the histologic and immunoperoxidase findings. Radioactivity in pelvic lymph nodes was prominently seen within 20 min of injection and was present 16 days later. Persistent accumulation of tracer in the lymphatics of the lower extremities was also observed in all patients 16 days post injection. Radioactivity counts in tumor-free lymph nodes were higher than in tumored lymph nodes resected. Our results demonstrate that intra lymphatic administration of 111In-labeled PAY 276 monoclonal antibody has major technical limitations, and that further research directed at the causes of tracer accumulation in the lymphatics and tumor-free lymph nodes is required.     

Intraperitoneal 131I- and 111In-791T/36 monoclonal antibody in recurrent ovarian cancer: imaging and biodistribution

An examination of the biokinetics and biodistribution of i.p. administered 131I- or 111In-labelled 791T/36 antibody (1 mg) has been carried out in five patients with stage III/IV ovarian cancer. Blood kinetics and urinary excretion of the radiolabels were assayed. Scintigraphy was performed immediately following administration and before and after peritoneal lavage at 48 h. Blood levels of both preparations rose over the first 20-40 h reaching 8-14% of the administered dose in the circulation and then declined (T1/2 of 40 h). Circulating radiolabel was still attached partially to antibody as shown by precipitation with anti-mouse IgG antiserum. The rapid appearance of radiolabel in the bloodstream meant that any tumour localization could be from circulating antibody rather than local infiltration. Interpretation of the images was difficult and the distribution of the tracer was different from that previously observed using i.v. administration of antibody. In some cases the images were confusing and the uptake of activity did not fit in with the clinical knowledge of the disease or the findings from laparoscopy. Tumour specimens resected at 4-5 days showed up to 0.02% of the dose g-1.     

Indium-111 T101 monoclonal antibody is superior to iodine-131 T101 in imaging of cutaneous T-cell lymphoma

We have reported that [111In]T101 is highly effective in the detection of cutaneous T-cell lymphoma (CTCL) in nodal and cutaneous (erythroderma and tumor) sites. This study compares the biodistribution of [131I]T101 (1 to 7.1 mg, 2 mCi) in four patients with CTCL; two of these patients also received [111In]T101 (1 mg, 5 mCi). There was rapid clearance of [131I]T101 from whole-body, spleen, liver, and bone marrow, with evidence of loss of 131I tracer from the T101. Lymph node uptake was minimal in three of four patients, and there was no localization in skin lesions. This contrasted with [111In]T101 where there was prolonged retention of activity in these organs and excellent uptake in skin tumors, erythroderma, and lymph nodes. The study showed that [131I]T101 was suboptimal for imaging CTCL patients and demonstrates that the isotope or labeling method can dramatically alter the apparent biodistribution and tumor targeting of a given monoclonal antibody.     

Indium-111 antimyosin monoclonal antibody Fab imaging in patients with cardiomyopathy

Six patients with cardiomyopathy were imaged following intravenous injection of an indium-111 labeled monoclonal antibody directed against the heavy chain of cardiac myosin. Two patients had hypertrophic non-obstructive cardiomyopathy (HNCM), two patients had dilated cardiomyopathy (DCM), and two patients had specific heart muscle disease. One of 2 patients with HNCM and one of 2 patients with DCM had a positive antimyosin scan. The 2 patients with specific heart muscle disease manifested persistent blood pool activity of the antibody, thereby precluding interpretation of the images. The present report demonstrates that antimyosin antibody imaging may provide evidence of myocardial injury, or necrosis in some patients with cardiomyopathy.     

Thrombus imaging with indium-111 and iodine-131-labeled fibrin-specific monoclonal antibody and its F(ab')2 and Fab fragments

We have previously reported successful imaging of fresh (2-4 hr old) and aged (1-5 days old) canine thrombi with 131I-labeled intact monoclonal antibody (MAb) specific for fibrin. We now report thrombus imaging with 131I-labeled F(ab')2 and Fab and 111In-labeled intact MAb, F(ab')2, and Fab. Indium-111-labeled F(ab')2 proved to be the best imaging agent due to less nonspecific binding in the liver than whole IgG. Image quality was improved by the higher administered dose permissible with 111In and its better physical characteristics for imaging, compared to 131I. Immunofluorescence of fresh human histologic sections showed intact MAb and F(ab')2 binding to thrombi, pulmonary emboli, and atherosclerotic plaques, strengthening the feasibility of clinical thrombus imaging.     

Comparison of 99Tcm-labelled monoclonal anti-granulocyte antibody and 111In-labelled IgG for the detection of focal sites of infection in rats

The abilities of 99Tcm-labelled monoclonal anti-human granulocyte antibody (AGAb) and 111In-labelled nonspecific polyclonal human immunoglobulin (IgG) to localize at focal sites of inflammation were compared in rats with deep thigh infection due to E. coli. The radiolabelled antibodies were coadministered followed 4-6 and 24 h later by imaging and biodistribution studies. At 4-6 h after injection, the target to background ratio (T/B, lesion to contralateral leg) and percentage residual activity (% RA, counts in the lesion/total body counts) were nearly identical for both antibody preparations. At 24 h, T/B and % RA increased significantly (P less than 0.001) for both proteins but differences between the agents were not significant. In vitro analysis of the binding of AGAb and human polyclonal IgG to rat granulocytes showed a low level of binding with both agents. These results suggest that the primary mechanism of localization, by either antibody preparation in this model, is not antigen related. 111In-labelled nonspecific human IgG and 99Tcm-AGAb are equivalent reagents for the detection of focal sites of infection in the rat.     

Iodine-131 and indium-111 labelled avidin and streptavidin for pre-targetted immunoscintigraphy with biotinylated anti-tumour monoclonal antibody

Studies have been carried out in mice to examine by gamma camera imaging and dissection analysis the biodistribution of 131I- and 111In-labelled avidin and streptavidin. This has included mice with human tumour xenografts pre-targetted with biotinylated anti-tumour monoclonal antibody. Both iodine-labelled avidin and streptavidin were cleared rapidly from the circulation particularly to kidneys, which showed prolonged retention of the tracer. Similar distribution was seen with 111In-labelled preparations, although here tracer also accumulated in the liver. Radiolabelled preparations could form complexes in vitro with a biotinylated monoclonal antibody. This biotinylated antibody, following radioiodine labelling, localized in xenografts of a human osteosarcoma. When xenografted mice were injected with biotinylated antibody followed by 111In-labelled avidin, levels of tracer per gram of tumour tissue were four times higher than those in non-pretreated mice. However, there was still marked liver and kidney uptake of tracer and, since this dominated the images, tumours were not visualized by scintigraphy.