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     

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.     

Technetium-99m human polyclonal immunoglobulin G studies and conventional bone scans to detect active joint inflammation in chronic rheumatoid arthritis.

Rheumatoid arthritis is a chronic polyarthritis in which active inflamed joints coexist with joints in remission. We performed bone scans (99mTc-DPD) and 99mTc human polyclonal immunoglobulin G scans (99mTc-IgG) in 18 patients with rheumatoid arthritis to assess the uptake in actively inflamed joints and in joints in which remission after inflammation had occurred. A quantitative analysis of tracer uptake in each joint was performed on both scans. In 123 joints without current active inflammation, an increased 99mTc-DPD uptake was observed (2.31 +/- 1.27), whereas no 99mTc-IgG uptake was noted (1.18 +/- 0.32). Some 78 joints with mild pain or swelling exhibited increased 99mTc-DPD uptake (2.48 +/- 1.14) and increased 99mTc-IgG uptake (1.76 +/- 0.50; P less than 0.001), while 21 joints with moderate to severe pain or swelling exhibited increased 99mTc-DPD uptake (2.39 +/- 0.93) and increased 99mTc-IgG uptake (1.79 +/- 0.51; P less than 0.001). In conclusion, 99mTc-IgG scans distinguish between joints with and without active inflammation in chronic rheumatoid arthritis, whereas bone scans do not. Thus, 99mTc-IgG scans may be useful in identifying joints with current active inflammation in rheumatoid arthritis.     

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).     

Measurement of lymphatic function with technetium-99m-labelled polyclonal immunoglobulin.

A reliable method for measuring lymph flow in physiological units would be valuable, especially in conditions in which it is uncertain whether lymph flow is increased or decreased. The requirements of a radiopharmaceutical for such measurement include stable radionuclide labelling and rapid access to lymphatic vessels following tissue injection but no access to blood vessels. A soluble macromolecule is likely to come closest to meeting these requirements. Technetium-99m-labelled human polyclonal immunoglobulin (HIG) was therefore investigated firstly in comparison with 99mTc-labelled human serum albumin (HSA) in patients undergoing routine lymphoscintigraphy and secondly with respect to injection site in a group of volunteers with post-mastectomy oedema (PMO). Subcutaneous injection of 99mTc-HIG into the web space of a distal extremity gave images in which lymphatic vessels were more clearly defined compared with images obtained after injection of 99mTc-HSA. Lymph nodes were also more clearly defined, suggesting specific retention of HIG, possibly through Fc-mediated binding. Peripheral blood sampling showed a delayed arrival in blood of radioactivity after 99mTc-HIG compared with 99mTc-HSA, although ultimately, the blood recovery of 99mTc-HIG was significantly higher (P < 0.05) than that of 99mTc-HSA. Clearance rates of radioactivity from the injection site were not significantly different, however, between the two agents. In patients with PMO, web space injection of 99mTc-HIG gave excellent images of normal lymphatic vessels, of lymph nodes and of abnormal lymph drainage such as dermal backflow in swollen arms. In contrast, neither lymphatic vessels nor lymph nodes were visualised after injection into the skin of the dorsum of the distal forearm. Although there was no difference in clearance rates from the injection sites between normal and swollen arms with either agent in PMO, clearance was significantly faster following injection into the web space (0.11% per minute for normal and swollen arms combined) than into the forearm (0.053% per minute; P < 0.05). These results suggest that (a) 99mTc-HIG is a potentially useful agent for measuring lymph flow and lymph node function; but (b) injection into the dorsum of the forearm is not a useful method of administration for these measurements; and (c) clearance rates from the injection site do not support the notion that PMO is the result of decreased lymph flow. Further studies are warranted to evaluate 99mTc-HIG as an agent for assessment of lymphatic function, especially with respect to measurement of lymph flow and possibly also for the evaluation of lymph node Fc-mediated immunocompetence.     

Technetium-99m labelled hydrazinonicotinamido human non-specific polyclonal immunoglobulin G for detection of infectious foci: a comparison with two other technetium-labelled immunoglobulin preparations

Recently a new linker — hydrazinonicotinate (HYNIC) — was introduced for labelling of proteins and peptides with technetium-99m. HYNIC and other linkers have been used for labelling of human non-specific polyclonal immunoglobulin G (hIgG) with^99mTc for the detection of infections. In this study we compared the tissue distribution of three different^99mTc-hIgG preparations in groups of five Wistar rats with a focal intramuscular infection withStaphylococcus aureus. We compared^99mTc-HYNIC-hIgG with^99mTc-hIgG labelled via the so-called Schwarz method (reduction of disulphide bonds) and with the^99mTc-labelled commercially available Technescan-HIG. Unlike the HYNIC linker, in the two other labelling methods free sulph-hydryl groups are involved in the binding of^99mTc. High-performance liquid chromatography analysis of the labelled preparations and of plasma samples revealed aggregate or polymer formation in all three agents; this was least pronounced in the product labelled by means of the Schwarz method. The tested preparations did not show signs of degradation in vitro. The difference in linker chemistry was reflected in the tissue distribution. Thus the biodistribution of^99mTc-HYNIC-hIgG was significantly different from the distribution of the two other preparations: abscess (1.4%±0.2%ID/g), muscle, liver, spleen, plasma, lung, bone marrow, and small intestine concentrations were higher at 24 h p.i.; kidney uptake (1.19%±0.08%ID/g) was significantly lower. The abscess-to-plasma and the abscess-to-muscle ratios (0.5 and 11, respectively), however, were in the same range for the three preparations tested. Quantitative analysis of the scintigraphs revealed that the total body clearance of^99mTc-HYNIC-hIgG was significantly slower than for the other agents. The abscess uptake of^99mTc-HYNIC-hIgG as a percentage of the remaining body activity was significantly higher. Based on its high abscess uptake, its low uptake in the kidneys and the high percentage of its abscess uptake in relation to the remaining body activity, we conclude that^99mTc-HYNIC-hIgG seems superior to the two other preparations tested for the detection of infections.     

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.     

Tc-99m polyclonal human immunoglobulin G imaging in Graves' ophthalmopathy

PURPOSE: The authors evaluated the utility of Tc-99m-labeled human immunglobulin G (HIG) in determining the severity of orbital inflammation and the relation of orbital Tc-99m HIG uptake and clinical parameters in patients with Graves' ophthalmopathy. MATERIALS AND METHODS: Images were obtained in 23 patients (13 women, 10 men; mean age, 51+/-10 years) with Graves' ophthalmopathy. Planar orbital images were obtained and SPECT was performed using a triple-detector gamma camera 4 hours after 370 MBq (10 mCi) Tc-99m HIG injection. Tc-99m HIG uptake was classified using transaxial and coronal slices as 1, mild; 2, moderate; and 3, severe. The clinical severity of orbital disease was categorized, according to the criteria described by Feldon and Unsold, as class I, mild involvement; class II, moderate; and class III, severe. Disease was considered to be clinically inactive if symptoms and signs were stable or improved in the last two examinations performed at least 6 months apart. RESULTS: Sixteen patients were clinically inactive, and seven patients were active. The mean Tc-99m HIG classes were 1.5+/-0.5 and 2.6+/-0.5, respectively (P = 0.02). There was not a good correlation between the clinical classification and Tc-99m HIG classification, whereas the presence of active disease showed a good correlation with Tc-99m HIG classification (r = 0.703; P = 0.0002). CONCLUSIONS: Tc-99m HIG imaging showed possible ongoing subclinical inflammation in the orbits of the patients with Graves' ophthalmopathy regardless of the clinical classification. Tc-99m HIG SPECT seems a promising procedure for evaluating the presence of active orbital inflammation.     

Technetium-99m human polyclonal immunoglobulin G studies and conventional bone scans to detect active joint inflammation in chronic rheumatoid arthritis

Rheumatoid arthritis is a chronic polyarthritis in which active inflammed joints coexist with joints in remission. We performed bone scans (^99mTc-DPD) and ^99mTc human polyclonal immunoglobulin G scans (^99mTc-IgG) in 18 patients with rheumatoid arthritis to assess the uptake in actively inflamed joints and in joints in which remission after inflammation had occurred. A quantitative analysis of tracer uptake in each joint was performed on both scans. In 123 joints without current active inflammation, an increased ^99mTc-DPD uptake was observed (2.31 ± 1.27), whereas no ^99mTc-IgG uptake was noted (1.18±0.32). Some 78 joints with mild pain or swelling exhibited increased ^99mTc-DPD uptake (2.48 ± 1.14) and increased ^99mTc-IgG uptake (1.76 ± 0.50; P <0.001), while 21 joints with moderate to severe pain or swelling exhibited increased ^99mTc-DPD uptake (2.39±0.93) and increased ^99mTc-IgG uptake (1.79±0.51; P <0.001). In conclusion, ^99mTc-IgG scans distinguish between joints with and without active inflammation in chronic rheumatoid arthritis, whereas bone scans do not. Thus, ^99mTc-IgG scans may be useful in identifying joints with current active inflammation in rheumatoid arthritis.Work was supported by a research grant from Mallinckrodt Ibér-ica Offprint requests to: L. Berná     

Contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin at sites of inflammation

The purpose of this study was to assess the contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin (HIG) at sites of inflammation. Mice were intraperitoneally injected withStaphylococcus aureus (SA animals), with heat-inactivated newborn calf serum (NBCS, to mimic a non-bacterial inflammation) or with physiological saline (controls); 1 h thereafter they received HIG. At various intervals after the administration of HIG the mice were killed, and the percentages of radioactivity in the peritoneal effluent and attached to the cellular and bacterial fraction thereof were established. Furthermore, the total number of cells and that of bacteria in the fluid were quantitated. The percentage of activity in the effluent in the SA animals was (P<0.02) higher than those in the NBCS-injected animals and controls from 4 h onwards. In all groups of mice this percentage was highest at 4 h and decreased (P<0.01) afterwards. The percentage of cell-bound activity and the total number of cells remained fairly constant or increased with time in the SA animals (P<0.01). The bacteria-bound activity remained rather constant throughout the experiment and ranged between 4% and 6%. In the SA-infected animals the percentage of cell-bound activity was correlated with the total number of cells (macrophages but especially neutrophils) but even more strongly with the number of cell-associated bacteria. In the NBCS-injected animals a correlation was demonstrated between the cell-bound activity and the total number of cells (only neutrophils). It is concluded that in both experimental inflammations, phagocytic cells, and especially neutrophils, contributed significantly to the accumulation of label at the site of inflammation. Their impact on this localization is augmented by the phagocytosis of bacteria.     

Detection of acute inflammation with In-Labeled nonspecific polyclonal IgG

The detection of focal sites of inflammation is an integral part of the clinical evaluation of the febrile patient. When anatomically distinct abscesses are present, lesion detection can be accomplished by standard radiographic techniques, particularly in patients with normal anatomy. At the phlegmon stage, however, and in patients who have undergone surgery, these techniques are considerably less effective. While radionuclide methods, such as Gallium-67 (67Ga)-citrate and Indium-111 (111In)-labeled WBCs have been relatively successful for the detection of early inflammation, neither approach is ideal. In the course of studies addressing the use of specific organism-directed antibodies for imaging experimental infections in animals, we observed that nonspecific polyclonal immunoglobulin G (IgG) localized as well as specific antibodies. Preliminary experiments suggested that the Fc portion of IgG is necessary for effective inflammation localization. Since polyclonal IgG in gram quantities has been safely used for therapy in patients with immune deficiency states, we decided to test whether milligram quantities of radiolabeled IgG could image focal sites of inflammation in humans. Thus far, we have studied a series of 84 patients with suspected lesions in the abdomen, pelvis, vascular grafts, lungs, or bones/joints. In 48 of 52 patients with focal lesions detected by surgery, computed tomography (CT), magnetic resonance imaging (MRI), or ultrasound (US), the IgG scan correctly localized the site, while 31 patients without focal inflammation had no abnormal focal localization of the radiopharmaceutical. Four patients had false negative scans and one patient had a false positive scan. For this small series, the overall sensitivity and specificity were 92% and 95%, respectively. In this report, we review our experience with this exciting new agent.     

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.     

Measurement of lymphatic function with technetium-99m-labelled polyclonal immunoglobulin

A reliable method for measuring lymph flow in physiological units would be valuable, especially in conditions in which it is uncertain whether lymph flow is increased or decreased. The requirements of a radiopharmaceutical for such measurement include stable radionuclide labelling and rapid access to lymphatic vessels following tissue injection but no access to blood vessels. A soluble macromolecule is likely to come closest to meeting these requirements. Technetium-99m- labelled human polyclonal immunoglobulin (HIG) was therefore investigated firstly in comparison with ^99mTc-labelled human serum albumin (HSA) in patients undergoing routine lymphoscintigraphy and secondly with respect to injection site in a group of volunteers with post-mastectomy oedema (PMO). Subcutaneous injection of ^99mTc-HIG into the web space of a distal extremity gave images in which lymphatic vessels were more clearly defined compared with images obtained after injection of ^99mTc-HSA. Lymph nodes were also more clearly defined, suggesting specific retention of HIG, possibly through Fc-mediated binding. Peripheral blood sampling showed a delayed arrival in blood of radioactivity after ^99mTc-HIG compared with ^99mTc-HSA, although ultimately, the blood recovery of ^99mTc-HIG was significantly higher (P <0.05) than that of ^99mTc-HSA. Clearance rates of radioactivity from the injection site were not sinificantly different, however, between the two agents. In patients with PMO, web space injection of ^99mTc-HIG gave excellent images of normal lymphatic vessels, of lymph nodes and of abnormal lymph drainage such as dermal backflow in swollen arms. In contrast, neither lymphatic vessels nor lymph nodes were visualised after injection into the skin of the dorsum of the distal forearm. Although there was no difference in clearance rates from the injection sites between normal and swollen arms with either agent in PMO, clearance was significantly faster following injection into the web space (0.11% per minute for normal and swollen arms combined) than into the forearm (0.053% per minute; P<0.05). These results suggest that (a) ^99mTc-HIG is a potentially useful agent for measuring lymph flow and lymph node function; but (b) injection into the dorsum of the forearm is not a useful method of administration for these measurements; and (c) clearance rates from the injection site do not support the notion that PMO is the result of decreased lymph flow. Further studies are warranted to evaluate ^99mTc-HIG as an agent for assessment of lymphatic function, especially with respect to measurement of lymph flow and possibly also for the evaluation of lymph node Fc-mediated immunocompetence. Received 28 July and in revised form 26 October 1998     

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.     

A prospective comparison of 99mTc-labeled polyclonal human immunoglobulin and 111In granulocytes for localization of inflammatory bowel disease.

There is a need for an easily prepared radiopharmaceutical agent for the detection of inflammation and infection. In a group of 14 patients with inflammatory bowel disease (IBD), the detection of actively involved intestinal segments by nonspecific human polyclonal immunoglobulin (IgG) labeled with 99mTc was compared with that of 111In granulocytes. To determine the specificity of 99mTc-IgG scintigraphy, 8 control patients without clinical indications of intestinal inflammation were examined. 99mTc-IgG was found in the left colon in 8 and in the right colon in 7 of the 8 controls 4 hours after the injection. At that time of scintigraphy only 4 IBD patients exhibited a more intense accumulation at the site of the intestinal segments with active disease. In contrast, in a randomized comparison with 111In granulocytes scintigraphy was positive in 11 patients with the latter technique. Moreover, fewer diseased segments were seen in the 4 patients with positive 99mTc-IgG scintigraphy (6 versus 12 with 111In granulocytes). In view of the low sensitivity and specificity, it is concluded that 99mTc-IgG is not suitable for the scintigraphic staging of IBD patients.