Localization of Fc and Fab fragments of nonspecific polyclonal IgG at focal sites of inflammation

Intact IgG, Fc, Fab, and 1/2Fc (reduced and alkylated Fc) were coupled to DTPA, labeled with indium-111 and administered to rats to compare the ability of fragments of IgG to localize at focal sites of inflammation. Two sets of experiments were performed: First, 1, 6, 24, and 48 hr after injection, biodistribution was determined in healthy animals; second, localization at sites of inflammation was determined by scintillation camera imaging of animals with deep-thigh infection due to Escherichia coli. The biodistribution studies demonstrated differences in kidney and liver localization: IgG less than Fc less than Fab less than 1/2Fc (kidney), Fc less than 1/2Fc less than IgG less than Fab (liver). The imaging studies revealed that target-to-background ratio (T/B) and percent residual activity (%RA) for IgG was significantly greater (p less than 0.01) than 1/2 Fc or Fab, and T/B for IgG was greater (p less than 0.01) than Fc. These studies suggest that the Fc portion of IgG is the fragment with the best imaging properties.     

Comparison of indium-111 nonspecific polyclonal IgG with indium-111-leukocytes in a canine osteomyelitis model

Osteomyelitis was surgically produced in the proximal tibia of ten dogs. A sham operation was performed on the other tibia. Early (3 hr) and late (20 hr) imaging was performed 1, 4, 7, 10, and 13 wk later, while the osteomyelitis progressed from acute to chronic. Indium-111-IgG had a significantly greater accumulation at the osteomyelitis site than 111In-leukocytes, both during early (p = 0.001) and late (p = 0.03) imaging, and at each of the weeks studied (p less than 0.001). During early imaging, both agents gave equivalent lesion to background ratios. On the late images, the 111In-leukocytes gave significantly higher lesion-to-background ratios than 111In-IgG (p less than 0.001) and higher ratios than they did during the early images (p less than 0.001). Both agents had greater accumulation in acute osteomyelitis than in chronic osteomyelitis (p less than 0.02). Osteomyelitis in the surgical site can be distinguished from the uptake in the sham surgery site using 111In-leukocytes, but not when using 111In-IgG.     

Radionuclide imaging of experimental atherosclerosis with nonspecific polyclonal immunoglobulin G

The utility of nonspecific polyclonal IgG for external imaging of experimental atherosclerosis was tested in a series of rabbits after balloon catheter deendothelialization of the abdominal aorta. Following injection of 111In-IgG, 111In-Fc, or 111In-Fab serial images were recorded. In addition, several animals received 125I-low density lipoproteins [125I-LDL], or 125I human serum albumin [125I-HSA] as positive and negative controls. Forty-eight hours after injection of the radiolabeled proteins, the aortas were removed, divided into abdominal and thoracic regions, counted, and autoradiographed. The images acquired after injection of 111In-IgG and 111In-Fc, showed clear focal accumulation of radioactivity in the healing abdominal aorta. In contrast, the images obtained after injection of 111In-Fab did not show focal radionuclide accumulation. For 111In-IgG and 111In-Fc there were three to six times as many counts in the abdominal as in the thoracic aorta, while for 111In-Fab and 125I HSA, the abdominal and thoracic counts were nearly equal. The results suggest that radiolabeled IgG and Fc can be used to image experimental atherosclerosis.     

Infectious imaging with indium-111-labeled nonspecific polyclonal human immunoglobulin

Nonspecific polyclonal immunoglobulin (IgG), prepared from pooled human serum gamma globulin and labeled with 111In has been reported to be equivalent to antigen-specific antibody in the detection of focal infection or inflammation during the first 24 hr after injection. We describe our experience in a Phase II clinical study using 111In-IgG in 15 patients (8 males, 7 females) ranging from 26 to 80 (mean = 50) yr of age with suspected focal infection/inflammation. Pathologic confirmation was obtained in 5/15 cases. A combination of clinical course, laboratory results, and other imaging procedures were used to categorize the other 10 patients. One possible false-negative involved a presumed aspiration pneumonia in a patient with a history of aspiration, bibasilar infiltrates on chest film, and no other identified source of infection. Otherwise, there were 10 confirmed positives, 4 confirmed negatives, and no false-positives. Our findings confirm earlier reports that 111In-IgG may be a superior imaging agent for infection/inflammation with practical advantages over 67Ga-citrate and 111In-labeled leukocytes.     

Radiolabeled, nonspecific, polyclonal human immunoglobulin in the detection of focal inflammation by scintigraphy: comparison with gallium-67 citrate and technetium-99m-labeled albumin

The accumulation of nonspecific polyclonal human immunoglobulin (IgG) radiolabeled with 125I or 111In was compared to that of [67Ga]citrate and [99mTc]albumin in rats with deep thigh inflammation due to Escherichia coli infection. Serial scintigrams were acquired at 1, 3, 24, and in some cases, 48 hr after injection. As early as 3 hr postinjection, [111In]IgG showed greater accumulation at the lesion than [99mTc]HSA (p less than 0.01). Both [125I]IgG and [111In]IgG showed greater accumulation than [67Ga]citrate (p less than 0.01). At 24 hr, IgG image definition increased, while HSA image definition decreased, and the intensity of accumulation of both IgG preparations was greater than that of [67Ga]citrate or [99mTc]HSA (p less than 0.01). At all imaging times, [67Ga]citrate accumulation was surprisingly low. In inflammation produced by Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Candida albicans, or turpentine, [111In]IgG accumulation was similar to the results obtained with Escherichia coli. These studies suggest that focal sites of inflammation can be detected with radiolabeled nonspecific human polyclonal IgG.     

Scintigraphic detection of bone and joint infections with indium-111-labeled nonspecific polyclonal human immunoglobulin G

The utility of indium-111-(111In) labeled immunoglobulin G (IgG) to detect infection of bone and adjacent tissues was investigated. Proof of infection was obtained by cultures taken at surgery. All 32 patients showed focally increased uptake on the technetium-99m- (99mTc) methylene diphosphonate (MDP) skeletal scintigraphies. Labeled immunoglobulin correctly identified presence, location, extent and soft-tissue involvement of the suspected inflammatory site. In these patients, focally increasing accumulation was noted over 48 hr. Discrimination between infection and sterile inflammatory lesions was not possible. Two fractures, 6-mo-old, and an aseptic loosening of a total-hip prosthesis were not visualized. Side effects after the immunoglobulin administration were not observed. Radiolabeled immunoglobulin is a new and safe radiopharmaceutical for the investigation of infectious bone and joint disease. The sensitivity of this agent appears at least as high as that of labeled leukocytes. However, labeled immunoglobulin can easily be prepared in every nuclear medicine department.     

Localization of infection using streptavidin and biotin: an alternative to nonspecific polyclonal immunoglobulin.

Since favorable images of infection are obtained with radio-labeled nonspecific IgG, streptavidin has been considered as an alternative protein in this investigation. The advantage of streptavidin is that once localized it may be targeted with radiolabeled biotin. Studies were conducted in a mouse model with an Escherichia coli infection in one thigh. Indium-111-labeled streptavidin showed equivalent localization to the infection as that obtained with 111In-labeled polyclonal nonspecific IgG, however blood levels with streptavidin were lower at all time points; consequently, target-to-blood ratios were improved. Pretargeting with unlabeled streptavidin followed 3 hr later with 111In-labeled biotin showed equivalent localization in the target and reduced activity in all organs sampled. As such, infected thigh-to-normal thigh ratios were improved 3-fold for pretargeting versus either labeled IgG or streptavidin. Improvements in infected thigh-to-liver and blood ratios were greater than 8-fold. Only in the case of kidneys was the ratio unimproved. In conclusion, we have shown that by preadministration of unlabeled streptavidin followed by labeled biotin, infectious lesions in a mouse model may be imaged earlier with lower background levels relative to the administration of labeled nonspecific IgG.     

Measurement of synovial inflammation in rheumatoid arthritis with technetium 99m labelled human polyclonal immunoglobulin G

The ability of technetium 99m labelled nonspecific, polyclonal human immunoglobulin G (99mTc-IgG) scintigraphy to depict and quantify synovial inflammation was studied in patients with rheumatoid arthritis (RA). Eight patients with clinically active synovitis were injected with 350 MBq 99mTc-IgG, and imaging took place 4 h later. This resulted in excellent images of inflamed synovium. Significant correlations were observed between individual joint uptake on the scan and scores for joint pain (n = 316, p less than 0.001), joint swelling (n = 300, p less than 0.001) or the average of pain and swelling (n = 300, p less than 0.001). These results suggest that 99mTc-IgG scintigraphy may provide an objective, non-invasive test to detect and measure synovitis.     

Measurement of synovial inflammation in rheumatoid arthritis with technetium 99m labelled human polyclonal immunoglobulin G

The ability of technetium 99m labelled nonspecific, polyclonal human immunoglobulin G (^99mTc-IgG) scintigraphy to depict and quantify synovial inflammation was studied in patients with rheumatoid arthritis (RA). Eight patients with clinically active synovitis were injected with 350 MBq ^99mTc-IgG, and imaging took place 4 h later. This resulted in excellent images of inflamed synovium. Significant correlations were observed between individual joint uptake on the scan and scores for joint pain (n=316, p<0.001), joint swelling (n = 300, p < 0.001) or the average of pain and swelling (n = 300, p < 0.001). These results suggest that ^99mTc-IgG scintigraphy may provide an objective, non-invasive test to detect and measure synovitis. Offprint requests to: P.A.H.M. van der Lubbe     

Imaging of Pneumocystis carinii pneumonia with 111In-labelled non-specific polyclonal IgG: an experimental study in rats.

The relative sensitivity of imaging with 67Ga-citrate (67Ga) and non-specific human polyclonal IgG radiolabelled with 111In (111In-IgG) for the detection of Pneumocystis carinii pneumonia (PCP) was determined in rats. Pneumocystis carinii pneumonia was induced by low protein diet and dexamethasone. The course of the disease was monitored by serial imaging with 111In-IgG and 67Ga. Diffuse accumulation of both radiopharmaceuticals was observed in the lungs of infected animals (infection was verified by histological examination of the lungs), however, accumulation of 111In-IgG was consistently higher. In rats with early PCP, 111In-IgG imaging revealed pulmonary accumulation in animals with normal chest radiographs and 67Ga scans. In animals successfully treated for PCP, decreased pulmonary accumulation of 111In-IgG coincided with histological improvements. Several animals developed superinfection with bacteria or fungi. These animals had striking focal accumulation of 111In-IgG, in addition to the pattern of generalized uptake. Gallium concentration in these animals did not show this focal accumulation. These observations suggest that 111In-IgG may be useful for detecting PCP and pulmonary abscesses in the immunocompromised host.     

单/多克隆抗体在肾移植急性排斥反应中的临床应用研究

目的：探讨应用抗淋巴细胞单克隆／多克隆抗体治疗肾移植术后急性排斥反应的效果、安全性和副反应等。方法：我中心施行同种异体肾移植术后对激素冲击治疗不敏感的急性排斥患者43例，分为两组，分别给予单克隆抗体OKT3和多克隆抗体ALG治疗，并对治疗效果、安全性和药物副反应等进行分析。结果：ALG治疗25例，排斥逆转16例，治愈率64％；OKT3组18例，排斥逆转15例，治愈率83．3％；两组有显著性差异（P〈0．05）。结论：单／多克隆抗体作为肾移植术后排斥反应的治疗用药，临床效果肯定．可使一部分患者避免因排斥反应丧失移植肾。但同时它们进入人体能引起首剂综合征等很多副反应，应当引起重视。OKT3治疗耐激素性急性排斥反应效果优于ALG。     

Effect of isoelectric point on biodistribution and inflammation imaging with indium-111-labelled IgG

Electrostatic effects play an important role in protein interactions and may alter the biodistribution of antibodies. To study the effect of molecular charge on the biodistribution and infection imaging properties of human polyclonal immunoglobulin G (IgG), its iso electric point was varied by changing the level of diethylene triamine penta-acetic acid (DTPA) substitution: 0.8, 0.9, 3.7, 5.1 and 5.9 DTPA/IgG. Biodistributions of the different IgG preparations were determined at 10 min, 1, 6, 24, and 48 h post injection in normal rats, and infection imaging properties were determined in rats withEscherichia coli thigh infections. The biodistribution was significantly affected by pl. The immunoglobulin preparations with 0.9 and 3.7 DTPA/IgG showed faster clearance from the circulation and generally lower accumulation in most organs. The images had a target-to-background ratio of approximately 1.3–2.3:1. These results suggest that even though targeting is not affected by the level of DTPA substitutions, preparations with 0.9 and 3.7 DTPA/IgG may be superior imaging agents because of reduced accumulation by background organs.     

Detection of a local staphylococcal infection in mice with technetium-99m-labeled polyclonal human immunoglobulin.

The purpose of this study was to investigate both the ability of 99mTc-labeled polyclonal human immunoglobulin (HIG) to localize an infection and the modes of action involved in this process. Mice, infected with Staphylococcus aureus ATCC 25923 in a thigh muscle, received HIG intravenously. Scintigrams were made 1, 4, and 24 hr later; subsequently the mice were killed and the activity in several organs and thighs was determined. The radiopharmaceutical demonstrated a time-dependent accumulation at the site of infection. It was found that vascular permeability or Fc binding alone could not account for the mode of action of HIG. Neither the origin of Ig (human versus murine) nor the total amount of protein (0.01-1.0 mg Ig per mouse) affected the target-to-background (T/B) ratios. Ratios were not different for leukocytopenic animals. A correlation (p less than 0.001) was demonstrated between the number of bacteria at the site of infection and the T/B ratio. This was also found after antibiotic treatment (p less than 0.02).     

手掌侧腱滑囊非特异性炎症的CT诊断

目的 :探讨手掌侧腱滑囊非特异性炎症的诊断与鉴别诊断。方法 :5例行螺旋CT平扫 ,2例行增强扫描并做多平面重建 (MPR) ,层厚 5～ 10mm ,层距 5～ 10mm。结果 :CT显示腕尺侧滑液囊滑膜增生 ,滑液囊增宽、增厚、延长 ,其内肌腱排列次序紊乱、分散、推移而无增粗改变。增宽延长的滑液囊自腕横韧带上 3cm至掌骨远段止 ,而桡侧滑液囊正常 ,MPR显示尤为清楚。结论 :本病为非特异性炎症引起滑膜增生 ,可能与外伤及过度劳作有关。需与化脓性、结核性滑囊炎、单纯滑膜囊肿、腱鞘囊肿鉴别     

Detection of inflammation in aortic aneurysms with indium 111-oxine-labeled leukocyte imaging

BACKGROUND: The exact cause of aortic aneurysms is not completely understood. Histologically, the atherosclerotic lesions present in an aneurysm contain numerous inflammatory cells. This finding represents active atherosclerosis, which can cause lesion expansion. In this study we investigated the role of scintigraphy in the evaluation of inflammation in aortic aneurysms. METHODS AND RESULTS: We performed imaging using indium 111-oxine--labeled leukocytes in 14 patients with aortic aneurysms (10 thoracic and 4 abdominal) diagnosed by computed tomography. Peripheral blood evidence of inflammation was assessed on the same day. In 8 patients who subsequently underwent graft replacement of the aneurysm, the excised specimen was examined for evidence of inflammatory infiltration and correlated with the scintigraphic findings. Scintigraphic accumulation of labeled leukocytes was present in 10 of the 14 patients. Although all patients had a small increase in the erythrocyte sedimentation rate, there was no significant difference in the erythrocyte sedimentation rate between patients with positive and negative scintigram results. In 5 of the 8 surgical patients with positive scintigram results, the resected specimens demonstrated numerous inflammatory cells in the adventitia of the aortic wall and atherosclerotic changes in the media. There was no correlation between the presence of periaortic inflammatory adhesions at the time of surgery and the scintigraphic results. CONCLUSIONS: The accumulation of In-111-oxine--labeled leukocytes is a potentially useful scintigraphic marker of inflammatory infiltration in aortic aneurysms.     

Radiation treatment of acute inflammation in mice

PURPOSE: Low-dose radiotherapy (RT) has often been used effectively for the treatment of a variety of benign diseases, particularly those with acute inflammatory features. Here we report findings on radiation treatment of acute inflammation using a murine carrageenin air pouch model. MATERIALS AND METHODS: Air pouches raised on the dorsal surface of mice were injected with lambda carrageenin and were irradiated 6 h later with doses ranging from 0-5 Gy. Treatment success was evaluated at various times thereafter by volume of exudate and number of inflammatory cells, and levels of inflammation-related cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1beta) and transforming growth factor beta-1 (TGFbeta-1), and expression of inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), cyclooxygenase-2 (COX-2) and inducible heat shock protein 70 (HSP70) as determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. RESULTS: Crude inflammatory parameters such as the amount of exudates and number of inflammatory cells remained largely unaffected by radiation or were even a slightly and transiently increased. However, the expression of iNOS was attenuated by radiation concomitant with an increase in the levels of HO-1 and HSP70. Cytokine levels varied with the radiation dose and the time point. CONCLUSIONS: Ionizing radiation, even at low doses, functionally modulates inflammatory cells. Our findings indicate possible mechanisms as to how low-dose radiation may exert anti-inflammatory effects and provide the first evidence that heat shock proteins may be involved in this response.