The redistribution of granulocytes following E. coli endotoxin induced sepsis.

Infusion of endotoxin elicits granulocytopenia followed by increased numbers of granulocytes in peripheral blood. The purpose of this study was to investigate the redistribution and sequestration of granulocytes in the tissues following E. coli endotoxin induced sepsis. From 16 rabbits granulocytes were isolated, labelled with Indium and reinjected intravenously. Eight rabbits received an infusion of E. coli endotoxin 2 micrograms kg-1 while eight received isotonic saline. The redistribution of granulocytes was imaged with a gamma camera and calculated with a connected computer before and 2 and 6 hours after infusion of endotoxin or saline. Serum cortisol and interleukin-1 beta were measured. In another seven rabbits, respiratory burst activity and degranulation of granulocytes were measured prior to and from 5 min to 6 hours after infusion of E. coli endotoxin 2 micrograms kg-1 BW. Following infusion of endotoxin, the number of granulocytes in peripheral blood decreased from 2.44 to 0.064 x 10 l-1 two hours later. Within 5 min after infusion the overall oxidative burst of the peripheral blood granulocytes was increased and the granularity had decreased. Serum cortisol and interleukin-1 beta increased significantly. The radioactivity of labelled cells in the bone marrow and spleen decreased to 83.1% and 91.6% of initial values. At the same time there was a transient sequestration of labelled granulocytes in the lungs reaching 117.6% of initial values. The radioactivity of the liver increased continuously to 118.4%. The results indicate that endotoxin induces an efflux in activated granulocytes from peripheral blood, bone marrow and spleen to the lungs and liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies against chromosomal beta-lactamase.

A murine monoclonal anti-chromosomal beta-lactamase antibody was developed and an immunoblotting technique was used to study the presence of serum and sputum antibodies against Pseudomonas aeruginosa chromosomal group 1 beta-lactamase in patients with cystic fibrosis (CF). The serum antibody response was studied with serum samples collected in 1992 from 56 CF patients in a cross-sectional study and with serum samples from 18 CF patients in a longitudinal study. Anti-beta-lactamase immunoglobulin G antibodies were present in all of the serum samples from the patients with chronic bronchopulmonary P. aeruginosa infection (CF + P) but in none of the CF patients with no or intermittent P. aeruginosa infection. Anti-beta-lactamase antibodies were present in serum from CF + P patients after six antipseudomonal courses (median) and correlated with infection with a beta-lactam-resistant strain of P. aeruginosa. The sputum antibody response and the beta-lactamase activity in sputum samples from 14 of the CF + P patients were also studied. beta-lactamase antibodies were present in 10 of these samples. P. aeruginosa strains isolated from these samples were partially derepressed, producing group 1 cephalosporinase. We found a wide range of chromosomal beta-lactamase activity in the sputum samples, with no correlation with basal or induced activity of beta-lactamase expression. The presence of anti-beta-lactamase antibodies in endobronchial sputum could be an important factor in the defense against the infection. On the other hand, immune complexes between the beta-lactamase and corresponding antibodies could play a role in the pathogenesis of bronchopulmonary injury in CF by mediating hyperimmune reactions.

Redistribution of granulocytes during adrenaline infusion and following administration of cortisol in healthy volunteers.

Major surgical procedures induce an endocrine metabolic stress response characterized by increased secretion of adrenaline and cortisol. Furthermore, surgical stress is accompanied by granulocytosis in peripheral blood. The granulocytosis may be due to increased adrenaline and cortisol secretion. The purpose of this study was to investigate the redistribution of granulocytes during adrenaline infusion and following administration of cortisol. Granulocytes were isolated from peripheral blood from eight healthy volunteers, labelled with indium-111-tropolene and reinjected. The distribution of granulocytes was imaged by using a gamma camera and calculated with an interfaced computer three times during a control period and three times during a corresponding hour af adrenaline infusion 0.05 micrograms kg b.w.-1 min-1. The distribution was then measured every second h for another 6-h control period and then for 6 h following administration of cortisol 3.6 mg kg b.w.-1. Infusion of adrenaline resulted in granulocytosis in peripheral blood and a reduction of radioactivity of the spleen to 83.0% of the initial value. The effect of adrenaline on the bone marrow was negligible. Cortisol administration was followed by granulocytosis and decreased radioactivity of both the spleen (81.5%) and the bone marrow (79.8%). It is concluded that the spleen is an active immunological organ as both adrenaline and cortisol induces efflux of granulocytes from the spleen. The cortisol induced efflux of granulocytes from the bone marrow explains that granulocytosis also occurs in splenectomized patients after major surgery.

[99mTc anti-carcinoembryonic antigen in colorectal cancer]. / 99mTc-anti-karcinoembryonalt antigen ved kolorektal cancer.

This investigation is a part of a phase three multicentre immunoscintigraphy study of a technetium-99m labelled monoclonal antibody (Mab), BW 431/26, directed against carcino-embryonic antigen (CEA). Ten patients with recently colonoscopically verified primary colorectal cancer were evaluated. In nine of ten patients colorectal cancers were detected by wholebody immunoscintigraphy and by single photon emission computerized tomography (SPECT). The ten patients were operated upon, and in nine cases immunohistochemical investigations were performed. Immunohistochemical investigation of nine tumour specimens show that CEA was present in much higher concentrations in the tumour than in normal intestinal epithelium. Active accumulation of the labelled Mab in tumours was demonstrated by scintimetry. There is a demand for new diagnostic techniques for early diagnosis of colo-rectal cancer. Immunoscintigraphy must however compete with other diagnostic possibilities. The development of immunoscintigraphic technique is proceeding very rapidly, and there is probably a future for the use of immunological methods in diagnosis and therapy of cancer.

Immunoscintigraphy of colon cancers with the human monoclonal antibody COU-1.

The human monoclonal immunoglobulin M (IgM) antibody, COU-1 is obtained from a human-human hybridoma, which is derived by fusion between a human B-lymphoblastoid cell line and lymphocytes obtained from mesenteric lymph nodes from a patient with colorectal cancer. COU-1 recognizes a 43 kilodaltons intracellularly located cytokeratin-like protein, strongly expressed by adenocarcinoma tissue as compared to normal tissues. In tumor-bearing nude mice, antibody COU-1 labeled with 125I has been shown to accumulate in human colon cancer grafts when compared to human melanoma grafts and the normal mouse tissues. The observed accumulation was sufficient to be detected externally by immunoscintigraphy. Antigen-binding fragments of the antibody were also prepared and were shown to accumulate in colon cancer grafts. Improved tumor to normal tissue ratio was seen with the half-monomeric fragment, and the time required was reduced. In the clinic, five patients with suspected colorectal cancer were given 2 mg of 131I-labeled COU-1. No adverse effects were detected in any of the patients. Planar images were obtained on days 1, 2, 3, 5, and 7 after administration. The best images were obtained on days 5 and 7. Tumors were localized by immunoscintigraphy in four of the patients. Of these patients, surgery revealed that three of them had primary colorectal cancers located in the cecum, the ascending colon, and the rectum, respectively, while one patient had a pancreatic cancer. The smallest lesion observed had a diameter of 3 cm. In one of the patients, otherwise undiagnosed multiple liver metastases were revealed by the immunoscintigraphy and confirmed at surgery. An x-ray of the colon performed on the fifth patient had shown a stricture in the descending colon suspected to be caused by cancer. The tumor scintigraphy showed no accumulation of the antibody. Surgery revealed that the stricture was caused by adherence and not cancer. These findings are encouraging for further studies of this human monoclonal antibody in cancer patients.

Redistribution of granulocytes in patients after major surgical stress.

Major surgery induces a stress response characterized by granulocytosis in peripheral blood, and increased secretion of adrenaline and cortisol. The purpose of this study was to investigate the redistribution of granulocytes in response to major surgical stress. Granulocytes were isolated from eight surgical patients and eight healthy volunteers, labelled with Indium-111-tropolone, and reinjected. The distribution of granulocytes was imaged with a gamma camera and calculated by an interfaced computer before surgery and at 2, 4 and 6 h after the end of surgery. The volunteers served as a control group. In the hours after surgery the radioactivity of the area around the surgical field increased to 410.7% of initial values, while the radioactivity of the spleen decreased to 77.5%. In conclusion, the spleen constitutes a readily mobilizable source of granulocytes. This in vivo model demonstrates pronounced postoperative efflux of granulocytes to the area around the surgical field within hours.

Tumor detection with 131I-labeled human monoclonal antibody COU-1 in patients with suspected colorectal carcinoma.

A major factor limiting the use of rodent monoclonal antibodies for diagnosis and therapy of cancer is the development of human anti-mouse immunoglobulin antibodies. Here we report a phase I/II immunodetection study of a human monoclonal antibody, COU-1, labeled with 131I. COU-1 is produced by a human-human hybridoma and recognizes a M(r) 43,000 cytokeratin-like protein strongly expressed by adenocarcinomas of the colon, breast, and ovary. Ten patients were given an i.v. infusion of 2 mg of antibody COU-1 labeled with 185 MBq of 131I. No adverse effects or toxicity were detected by conventional clinical tests nor by a complement activation assay for C3d. None of the patients developed antibodies against antibody COU-1 as determined by enzyme-linked immunosorbent assay and agglutination analysis. Tumor detection was successful in 7 of 9 cancer patients. The tenth patient proved to be a true negative. In several instances immunoscintigraphy gave additional or more correct information than conventional detection techniques. Tumors were most clearly outlined at days 5 and 7 after infusion. Primary colorectal carcinomas were detected by planar imaging in the cecum, ascending colon, and rectum with the smallest lesion measuring 3.0 cm in diameter. Immunoscintigraphy revealed multiple liver metastases in 1 of 3 patients. However, the livers of all 3 patients contained significantly more radioactivity (P < 0.005) than tumor-free livers of the other patients. Pharmacokinetics was evaluated in all patients. The clearance of 131I-labeled COU-1 from the circulation followed a triphasic pattern; an initial phase [t1/2 = 0.4 +/- 0.4 (SD) h] cleared 23% of the radioactivity followed by a rapid phase with a half-life of 13 +/- 3.8 h. The third phase (beta-phase) exhibited a half-life of 119 +/- 36 h, which is similar to the half-life reported for normal IgM. The human monoclonal antibody COU-1 directed against a predominantly intracellular cancer-associated antigen does not produce toxicity or induce antibody formation and seems to be a promising agent for detecting tumors with immunoscintigraphy.

Redistribution of lymphocytes after major surgical stress.

Major surgery evokes an endocrine stress response, characterized by increased serum cortisol, plasma adrenaline and noradrenaline. Furthermore, surgical stress is accompanied by lymphopenia and granulocytosis in peripheral blood. The changes in peripheral white blood cells have been demonstrated after surgery as well as after cortisol infusion. The aim of the present study was to investigate to which tissues/organs peripheral blood lymphocytes are redistributed after major surgery. From 20 rabbits lymphocytes were isolated from peripheral blood, labelled with indium-111-tropolene and reinjected intravenously into the rabbits. Ten of the rabbits underwent major surgery (upper laparatomy) during general anaesthesia, while the control group (n = 10) was anaesthetized without surgery. The endocrine stress response to surgery was measured as serum cortisol, plasma adrenaline and noradrenaline. The redistribution of lymphocytes was imaged with a gamma camera and calculated with a connected computer before, 2, 4, and 7 h after the skin incision. Compared to preoperative values, laparotomy resulted in an increase in serum cortisol from 116.6 to 461.9 nmol/l (mean) and a decrease in the fraction/percentage of lymphocytes in peripheral blood from 43.8% to 14.7% 7 h after surgery. Simultaneously, the activity of the heart and lungs together decreased to 76.1% of initial values, while the spleen activity was unaffected. The radioactivity of the lymphatic tissue increased to 137.8% and 134.7%, respectively, 4 and 7 h after the start of surgery. The results indicate that major surgery induces a redistribution of lymphocytes from peripheral blood to lymphatic tissue. It is suggested that the endocrine stress response may be of major importance.

The redistribution of lymphocytes during adrenaline infusion. An in vivo study with radiolabelled cells.

Adrenergic activation is known to occur in sepsis and after major surgery or trauma. An elevated serum concentration of adrenaline is followed by lymphocytosis in peripheral blood even in splenectomized patients. The purpose of the present study was to clarify the redistribution of lymphocytes in the tissues during adrenaline infusion. Lymphocytes were isolated from 24 rabbits, labelled with indium-111-tropolone and reinjected into the rabbits. The next day the rabbits were anaesthetized. Eight rabbits received 3 micrograms of adrenaline i.v. followed by 0.2 micrograms/min, eight received 300 micrograms of adrenaline i.v. followed by 20 micrograms/min, while eight received a saline infusion and served as a control group. The activity of labelled cells was imaged with a gamma camera and computer before, during and after adrenaline infusion. The activity of the spleen decreased to 90% and 94% of initial values during low and high doses of adrenaline. The activity of the bone marrow decreased to 91% and 96%, respectively, while the activity of the heart/lung and the liver increased to 107% and 106% with the high dose of adrenaline. In peripheral blood the lymphocytes increased 10%. It is concluded that lymphocytes are redistributed from spleen and bone marrow to peripheral blood, lungs and liver during adrenaline infusion in this animal model.

Redistribution of lymphocytes after cortisol administration.

Major surgical procedures awake an endocrine metabolic stress response characterized by increased secretion of cortisol and lymphopenia. The purpose of this study was to clarify to which tissues the lymphocytes are redistributed after cortisol administration. Lymphocytes were isolated from 16 rabbits, labelled with indium-111-tropolone and reinjected into the rabbits. Eight of the rabbits received 25 mg of cortisol intravenously (group I), while eight received isotonic saline (group II). The redistribution of lymphocytes was imaged with a gamma camera and calculated by a connected computer before and two, four, and seven h after cortisol or saline administration. The radioactivity of cells from the spleen and bone marrow decreased to 84% and 56% of the initial values seven h after cortisol administration. The activity of the lymphatic tissues increased to 121% of initial values. It is concluded that during cortisol-induced lymphopenia the lymphocytes are redistributed from peripheral blood, spleen and bone marrow to lymphatic tissue.

Tumour localization and pharmacokinetics of iodine-125 human monoclonal IgM antibody (COU-1) and its monomeric and half-monomeric fragments analysed in nude mice grafted with human tumour.

Human monoclonal IgM antibodies reactive with cancer-associated antigens may not have the optimal imaging capability due to their large size. Fragmentation of human IgM is less than straight-forward due to the loss of immunoreactivity. From the human monoclonal IgM antibody COU-1 we have prepared monomeric and half-monomeric fragments, which retain the ability to bind to colon cancer cells in vitro. The pharmacokinetics and tumour localization were evaluated in nude mice bearing human colon adenocarcinoma and human melanoma grafts. Faster clearance from the circulation was seen for the smaller half-monomeric fragment with a half-life (rapid phase/slow phase) of 2 h/16 h compared with the intact antibody, 4 h/25 h, and the monomeric fragment, 3 h/27 h. Intact COU-1 as well as the fragments accumulated in the colon tumour graft. Higher amounts of radioactivity were found in the colon tumour as compared to normal organs for intact COU-1 at days 4 and 6, for the monomeric fragment at day 4, and for the half-monomeric fragment at day 2 after injection. This investigation demonstrates the favourable biodistribution of the half monomeric COU-1 fragment. The fast clearance of this fragment resulted in a tumour-to-muscle ratio as high as 22 on day 2 after injection. Also, only this fragment gave a positive tumour-to-blood ratio. Normal IgM and its fragments were used as controls. Radioimmunoscintigraphy demonstrated the colon tumour discriminatory properties of each of the three iodine-labelled antibody preparations.(ABSTRACT TRUNCATED AT 250 WORDS)

Indium-111-granulocyte scintigraphy in brain abscess diagnosis: limitations and pitfalls.

The scintigrams and records of 28 patients referred for indium-111-granulocyte scintigraphy (111In-GS) because of a suspected brain abscess were studied retrospectively. The final diagnosis was brain abscess in 8 patients, brain tumor in 18 patients, and infarct and hematoma in 1 patient each. Five patients not on corticosteroid treatment showed intense focal 111In accumulation in abscesses, whereas an abscess patient receiving a high steroid dose showed no uptake. Two patients studied twice showed intense uptake in abscesses when not on steroid therapy or on a low dose, whereas no uptake was seen when they received high or medium doses. Weak or moderate 111In uptake was observed in nine tumors. Microscopically assessed degree of tumor granulocyte infiltration, vessel proliferation, and hemorrhage did not correlate with the outcome of 111In GS. Our results suggest that intense focal cerebral 111In uptake favors the abscess diagnosis. Abscesses may go undetected, however, in patients on high- or medium-dose steroid therapy.

Intestinal permeability of 51Cr-labelled ethylenediaminetetraacetic acid in patients with Crohn's disease and their healthy relatives.

An increased intestinal permeability has been proposed as an aetiologic factor in Crohn's disease. The 24-h urinary excretion of 100 muCi 51Cr-labelled ethylenediaminetetraacetic acid (EDTA) was used to test the permeability in 15 patients with Crohn's disease and in 20 healthy first-degree relatives, who are known to have a genetic predisposition to inflammatory bowel disease. Twenty-eight healthy persons not related to patients with inflammatory bowel disease served as control material. The 51Cr-EDTA excretion of the relatives (range, 0.98%-3.87%; median, 1.935%) was not significantly higher than that of the controls (range, 1.17%-3.26%; median, 1.950%), whereas patients with Crohn's disease had a significantly higher excretion (range, 1.64%-8.86%; median, 2.940%) than both the relatives and the controls. Among patients the increased excretion was found only if the small intestine was involved. We conclude that 1) as a group, patients with Crohn's disease in the small intestine have an increased intestinal permeability, in contrast to their healthy relatives, who have a normal permeability; 2) a considerable overlap of the results of the 51Cr-EDTA test was found between the groups studied, and the test is not suitable for evaluating individual patients; 3) our results do not support the hypothesis of an increase in intestinal permeability as an aetiologic factor in Crohn's disease.

57Co-bleomycin scintigraphy for the preoperative detection and staging of lung tumors.

Twenty-five consecutive patients with lung tumors were classified according to the presence of metastases by the use of 57Co-Bleomycin scintigraphy. Twenty-two of the tumors were visualized but metastatic spread to hilar lymph nodes was not detected. Reliable separation of central tumors with and without growth into neighbouring organs was not possible. Mediastinal uptake was found in eight cases but only a part of these could be verified by operation. The method can contribute to the evaluation of lung tumors and can be helpful to avoid some unnecessary thoracotomies.

Accumulation of indium-111-labeled granulocytes in malignant tumors.

In a retrospective study of 220 [111In]granulocyte scintigrams from 208 patients, 25 patients had malignant neoplasms. Among these, tumor uptake of 111In activity was observed in ten patients (intense activity in two patients with non-Hodgkin's lymphoma and colonic carcinoma, respectively; moderate uptake in a patient with non-Hodgkin's lymphoma, and in a patient with an ovarian carcinoma; weak activity in three patients with cerebral neoplasms; and activity within otherwise "cold" metastatic lesions of the liver in three patients). Microscopic investigation following specific granulocyte staining revealed the greatest extent of granulocyte infiltration in the tumors which took up 111In activity, emphasizing the importance of tumor granulocyte infiltration as the single most important factor underlying tumor accumulation of 111In activity during [111In]granulocyte scintigraphy.

Analysis of factors that may affect the speed of accumulation of 111In-labelled granulocytes at sites of inflammation.

The mechanisms governing the accumulation of granulocytes in inflammatory lesions are poorly understood. Using a sensitive method of sequential 111In-granulocyte scintigraphy, we recorded the speed of focal 111In-granulocyte accumulation in 70 patients with non-osseous inflammatory and infectious foci, with special reference to the influence exerted by the duration of disease, patient age, body temperature, antibiotic therapy and initial trapping of granulocytes in the lungs. About 50% of the images had turned positive at 30 min after injection. Except for patients with urinary tract infections, the age of the patient did not influence the speed of 111In-granulocyte accumulation; nor did the duration of disease, antibiotic therapy or degree of initial granulocyte hold-up in the lungs. High fever, on the other hand, presumably reflecting an intense inflammatory reaction, was associated with an accelerated focal 111In-granulocyte accumulation, indicating that properties of the inflammatory process per se are major determinants of the speed of accumulation of 111In-labelled granulocytes in inflammatory processes.

Speed of accumulation of 111In-labelled granulocytes in focal non-osseous inflammatory processes.

Using a method of 111In-oxine granulocyte labelling in diluted plasma, we performed 162 scintigraphic studies in 159 patients with suspected non-osseous infection. We obtained a positive predictive value of 82%, and a negative predictive value of 97%, i.e. the method is very sensitive, albeit less specific. Sequential imaging showed around 50% of the scintigrams to turn positive at 30 min after the injection, the most rapid accumulation being seen in cases of superficial soft tissue infections and in pulmonary and pleural infections, cerebral abscesses showing a rather sluggish accretion of activity, urinary tract infections, abdominal abscesses and bowel inflammation accumulating activity at an intermediate rate. Our results indicate that the described method is sensitive, and that the sequential scintigraphic approach allows an early diagnosis in most cases, and facilitates the interpretation of delayed scans.