Radiolabelled leucocytes in human pulmonary disease.

Introduction: Radionuclides for leucocyte kinetic studies have progressed from non-gamma emitting cell-labelling radionuclides through gamma emitting nuclides that allow imaging of leucocyte kinetics, to the next goal of positron emission tomography (PET). Sources of data: Mostly the authors' own studies, following on from studies of the early pioneers. Areas of controversy: From early imaging studies, it appeared that the majority of the marginated granulocyte pool was located in the lungs. However, later work disputed this by demonstrating the exquisite sensitivity of granulocytes to ex vivo isolation and labelling, and that excessive lung activity is artefactual. Areas of agreement: Following refinement of labelling techniques, it was shown that the majority of marginated granulocytes are located in the spleen and bone marrow. The majority of leucocytes have a pulmonary vascular transit time only a few seconds longer than erythrocytes. The minority showing slow transit, ~5% in healthy persons, is increased in systemic inflammatory disorders that cause neutrophil priming and loss of deformability. Using a range of imaging techniques, including gamma camera imaging, whole-body counting and single photon-emission computerized tomography, labelled granulocytes were subsequently used to image pulmonary trafficking in lobar pneumonia, bronchiectasis, chronic obstructive pulmonary disease and adult respiratory distress syndrome. Growing points: More recently, eosinophils have been separated in pure form using magnetic bead technology for the study of eosinophil trafficking in asthma. Areas timely for developing research: These include advancement of eosinophil imaging, development of monocyte labelling, development of cell labelling with PET tracers and the tracking of lymphocytes.

Effects of tocilizumab on neutrophil function and kinetics.

BACKGROUND: Decreases in circulating neutrophils (polymorphonuclear leucocytes, PMNs) have been reported in patients treated with the anti-interleukin-6 receptor (IL-6R) antibody tocilizumab (TCZ); the mechanism for this is unclear. We hypothesize that TCZ reduces circulating neutrophils by affecting margination and/or bone marrow trafficking without affecting neutrophil function or apoptosis. MATERIALS AND METHODS: Eighteen healthy subjects were randomized to single intravenous dose of TCZ 8 mg/kg (n = 12) or placebo (n = 6) on day 0. On day 4, each subject had autologous indium-111-labelled neutrophils re-injected, and their kinetics quantified with longitudinal profiling in a whole body gamma-counter. TCZ-treated subjects were divided into two groups according to the extent of reduction in neutrophil count. RESULTS: Mean day 4 neutrophil counts, as % baseline, were 101·9%, 68·3% and 44·2% in the placebo, TCZ-PMN-'high' and TCZ-PMN-'low' groups, respectively (P < 0·001). Following TCZ, neutrophil function, activation and apoptosis ex vivo were all unaffected. In vivo, there were no differences in early blood recovery or margination to liver/spleen and bone marrow; however, later neutrophil re-distribution to bone marrow was markedly reduced in the TCZ-PMN-low group (peak pelvic count as % day 4 count on: day 5, 188% placebo vs. 127% TCZ-PMN-low, P < 0·001; day 10, 180% placebo vs. 132% TCZ-PMN-low, P < 0·01), with a trend towards higher liver/spleen neutrophil retention. CONCLUSIONS: We have demonstrated for the first time in humans that IL-6R blockade affects neutrophil trafficking to the bone marrow without influencing neutrophil functional capacity.

Use of 111-Indium-labeled autologous eosinophils to establish the in vivo kinetics of human eosinophils in healthy subjects.

Eosinophils are the major cellular effectors of allergic inflammation and represent an important therapeutic target. Although the genesis and activation of eosinophils have been extensively explored, little is known about their intravascular kinetics or physiological fate. This study was designed to determine the intravascular life span of eosinophils, their partitioning between circulating and marginated pools, and sites of disposal in healthy persons. Using autologous, minimally manipulated 111-Indium-labeled leukocytes with blood sampling, we measured the eosinophil intravascular residence time as 25.2 hours (compared with 10.3 hours for neutrophils) and demonstrated a substantial marginated eosinophil pool. γ camera imaging studies using purified eosinophils demonstrated initial retention in the lungs, with early redistribution to the liver and spleen, and evidence of recirculation from a hepatic pool. This work provides the first in vivo measurements of eosinophil kinetics in healthy volunteers and shows that 111-Indium-labeled eosinophils can be used to monitor the fate of eosinophils noninvasively.

Quantification of neutrophil migration into the lungs of patients with chronic obstructive pulmonary disease.

OBJECTIVE: To quantify neutrophil migration into the lungs of patients with chronic pulmonary obstructive disease (COPD). METHODS: Neutrophil loss via airways was assessed by dedicated whole-body counting 45 min, 24 h and 2, 4, 7 and 10 days after injection of very small activities of (111)In-labelled neutrophils in 12 healthy nonsmokers, 5 healthy smokers, 16 patients with COPD (of whom 7 were ex-smokers) and 10 patients with bronchiectasis. Lung accumulation of (99m)Tc-labelled neutrophils was assessed by sequential SPECT and Patlak analysis in six COPD patients and three healthy nonsmoking subjects. RESULTS: Whole body (111)In counts, expressed as percentages of 24 h counts, decreased in all subjects. Losses at 7 days (mean ± SD) were similar in healthy nonsmoking subjects (5.5 ± 1.5%), smoking subjects (6.5 ± 4.4%) and ex-smoking COPD patients (5.8 ± 1.5%). In contrast, currently smoking COPD patients showed higher losses (8.0 ± 3.0%) than healthy nonsmokers (p = 0.03). Two bronchiectatic patients lost 25% and 26%, indicating active disease; mean loss in the remaining eight was 6.9 ± 2.5%. The rate of accumulation of (99m)Tc-neutrophils in the lungs, determined by sequential SPECT, was increased in COPD patients (0.030-0.073 min(-1)) compared with healthy nonsmokers (0-0.002 min(-1); p = 0.02). CONCLUSION: In patients with COPD, sequential SPECT showed increased lung accumulation of (99m)Tc-labelled neutrophils, while whole-body counting demonstrated subsequent higher losses of (111)In-labelled neutrophils in patients who continued to smoke. Sequential SPECT as a means of quantifying neutrophil migration deserves further evaluation.

Measuring whole-body neutrophil redistribution using a dedicated whole-body counter and ultra-low doses of 111Indium.

BACKGROUND: There is increasing interest in the 'homing' of neutrophils to bone marrow. The aim of this study was to measure the whole-body redistribution of (111) In using a whole-body counter following the administration of ultra-small activities of (111) In-labelled neutrophils. METHODS: The detectors of a dedicated whole-body counter were fitted with lead collimators. Whole-body (111) In distribution was recorded at 45 min, 24 h, and 2, 4, 7 and 10 days after administration of (111) In-labelled neutrophils (0·29-0·74 MBq) in eight healthy non-smokers, five healthy smokers, eight patients with inactive bronchiectasis, three with asthma and nine with chronic obstructive pulmonary disease (COPD). RESULTS: Intravascular 45-min (111) In-labelled neutrophil recovery was not significantly different between groups, ranging from 33 (SD 8%) in healthy smokers to 45 (14%) in healthy non-smokers (P > 0·05). Peaks were identified on the whole body count profile corresponding to the chest, upper abdomen (liver/spleen) and pelvis (bone marrow). (111) In distribution changed between 45 min and 24 h and then remained stable thereafter. Peak chest counts increased ∼ 1·5-fold between 45 min and 24 h, whereas upper abdominal peak counts decreased by ∼ 25% with no significant inter-group differences. The increment in pelvic counts (∼ 2·7-fold) was similar between groups, except COPD patients, in whom it was 2·04 (0·35; P < 0·02 vs. healthy participants). CONCLUSIONS: Assuming neutrophils are distributed only between blood, liver, spleen and bone marrow, the data suggest that marrow pools 25% and destroys 67% of circulating neutrophils, rising in COPD to 40% and 80%, respectively, possibly as a result of the effects on marrow of chronic hypoxaemia.

Measurement of lymph node function from the extraction of immunoglobulin in lymph.

BACKGROUND: We aimed to measure the extraction fraction of human immunoglobulin G (HIG) by the 1st echelon lymph node (sentinel node) following intradermal injection in patients with breast cancer undergoing axillary lymph node dissection (ALND) and examine its association with node size and presence and extent of nodal metastatic disease. MATERIALS AND METHODS: HIG labelled with either In-111 (n = 21) or Tc-99m (n = 9) was injected intradermally at the areolar. ALND was performed 2-4 h later. All lymph nodes were isolated and individually counted in a well-counter. The counts in the 'hottest' (1st echelon) node were expressed as a fraction of total counts in all the resected nodes. Since counts in the least hot nodes barely exceeded background, this fraction represents extraction fraction for the 1st echelon node. Presence of disease was noted in each 1st echelon node and the extent quantified as percentage replacement with disease. RESULTS: Median extraction fraction in 1st echelon nodes with no or low (<1%) disease burden (n = 21) was 68 (range 23-93)%, significantly higher (p < 0.05) than in diseased 1st echelon nodes (n = 9), in which it was 44 (21-66)%. There was, however, no association between extraction fraction in diseased nodes and disease extent. In nodes with no/low disease, extraction fraction was similar for the two radiolabels. There was no association between extraction fraction and node size. CONCLUSION: Nodal extraction fraction of HIG is a novel physiological measurement. It is reduced as a result of metastatic invasion. In the absence of disease, it shows no correlation with node size.

Delivery of radiolabelled blood cells to lymphatic vessels by intradermal injection: a means of investigating lymphovenous communications in the upper limb.

OBJECTIVE: To identify peripheral lymphovenous communications (LVCs) using labelled erythrocytes and intradermal injection. Intradermal injection delivers macromolecules to loco-regional lymph nodes faster than subcutaneous injection, suggesting easier lymphatic vessel access. METHODS: Autologous erythrocytes labelled with 111In and 99mTc were injected into opposite hands. In four normal volunteers, the differentially labelled cells were given by intradermal injection on one side and subcutaneous injection on the other while in four breast cancer patients they were given by intradermal injection bilaterally 3 months after axillary lymph node clearance surgery. The axillae were imaged and blood samples obtained bilaterally at approximately 15, 30, 60, 120 and 180 min post-injection. Plasma activity was subtracted from whole blood activity to obtain erythrocyte-bound activity and contralateral concentrations were subtracted from ipsilateral concentrations to correct for ipsilateral recirculation. From estimated blood volume, erythrocyte and plasma activities contralateral to the injected side were calculated as percentage administered activity. Tracer concentrations in ipsilateral samples (%/l) were integrated to give total percentage administered activity, assuming a forearm blood flow of 20 ml/min. RESULTS: Kinetics of plasma activity were consistent with small diffusible 99mTc complexes and protein-bound 111In. With both radionuclides, axillary nodes were visualized after intradermal but not subcutaneous injection, suggesting that nodal activity arises from erythrocytes. In one patient, 99mTc and 111In labelled erythrocytes accumulated in similar amounts ipsilaterally and contralaterally, suggesting bilateral LVCs distal to the ipsilateral sampling point. There was no evidence of LVCs in the other seven volunteers. CONCLUSION: Intradermally injected erythrocytes are able to detect and potentially quantify peripheral LVCs.

Axillary lymph node drainage pathways from intradermal and intraparenchymal breast planes.

BACKGROUND: To compare functional anatomy of breast peri-areolar and peri-tumoral lymphatic drainage basins. METHODS: Fifteen breast cancer patients received simultaneous peri-areolar (intradermal) and peri-tumoral (intraparenchymal) injections of human polyclonal immunoglobulin (HIG) labeled with (99m)Tc and (111)In 2 to 4 h before axillary lymph node clearance surgery. Resected nodes (range 5-20; median 16) were individually counted for (99m)Tc and (111)In in a well-counter and ranked according to activity content (echelon). Activity in distal nodes was negligible so extraction efficiency (E) of HIG in the first echelon node was calculated as counts divided by total counts in the chain. RESULTS: Five- to 10-fold more activity was recovered after intradermal injection. The injection planes identified the same first echelon node in 10 patients (group 1) but different in five (group 2). In group 1, intradermal E correlated with intra-parenchymal E (r = 0.82; P < 0.01). E of intradermal first echelon nodes in group 2 was 51 (SD 13)%, similar to intradermal E in group 1 (58 [23]%). E of intraparenchymal first echelon nodes in group 2, however, was 28 (6)%, lower than intraparenchymal E in group 1 (54 [20]%; P < 0.02). CONCLUSIONS: Lymph nodes extract approximately 50% of HIG. Extracted HIG does not cascade to distal nodes, validating HIG for sentinel node lymphoscintigraphy. HIG injected intradermally at the areola drains via a single route to the axilla. In two-thirds of patients, peri-tumoral HIG follows a similar route, but in one-third of patients drainage from the parenchymal plane is more complex, with more than one route to the axilla.

Functional variation in lymph node arrangements within the axilla.

The aim of the project was to identify how lymphatic pathways are functionally arranged within the axilla (i.e., single linear chains, branching chains, and networks). We used ex vivo dual isotope radioassay of individual nodes resected at axillary lymphatic clearance surgery in breast cancer patients given simultaneous intradermal breast and intradermal hand injections (n = 15) or simultaneous intradermal breast and parenchymal breast injections (n = 15) of differentially labelled human immunoglobulin (Tc-99m-HIG and In-111-HIG). Nodes were ranked according to isotope content and activity-rank profiles constructed for each of the two injection sites. The majority of profiles following intradermal breast injection (17/30) were mono-exponential, consistent with a simple linear chain of nodes, with each node extracting a constant fraction of incoming HIG. In 15/17 of these, the accompanying profile from the alternative injection site was also mono-exponential and, in 11/15, essentially parallel. The profile appeared biphasic in 12/30 intradermal breast injections and of these 9/12 were accompanied by a biphasic profile (7/9 parallel) from the alternative injection site. In one patient, both profiles were polyphasic and parallel. Considering the respective shapes of paired profiles and whether the two injection sites shared the same first echelon nodes, functional lymph node arrangements are proposed. The commonest is a single linear chain, then a chain branching into two linear chains, and, least common, a network.

Lymphatic drainage pathways of the breast and the upper limb.

OBJECTIVE: To determine how often the sentinel lymph node (SLN) draining the breast is the same node as the SLN draining the upper limb. A common SLN might increase the risk of upper limb breast cancer-related lymphoedema after SLN biopsy. METHODS: Patients with invasive breast cancer, identified as being suitable for axillary lymph node dissection, were injected preoperatively with 40 MBq of technetium-99m (Tc)-human polyclonal immunoglobulin G intradermally into the ipsilateral breast and 3 MBq of indium-111 (In)-human polyclonal immunoglobulin G intradermally into the ipsilateral hand, or vice versa. Axillary lymph nodes were removed, separated and assayed in a well counter for Tc and In. RESULTS: Fifteen patients entered the study. In 13 of 15 patients, the 'hottest' lymph node for Tc was separate from the 'hottest' lymph node for In. In two of 15 patients the 'hottest' lymph node for Tc was also the 'hottest' lymph node for In, suggesting a common drainage pathway from the ipsilateral breast and upper limb. CONCLUSION: Although the majority of patients has different pathways of lymphatic drainage from the ipsilateral breast and upper limb, in a small minority of patients the drainage pathway is through a common SLN. Such patients may be at increased risk of developing upper limb breast cancer-related lymphoedema after SLN biopsy.

Utility of 111In-labelled leucocyte scintigraphy in patients with fever of unknown origin in an era of changing disease spectrum and investigational techniques.

BACKGROUND: (111)In-labelled leucocyte, imaging is often used to investigate patients with fever of unknown origin (FUO). Its diagnostic performance, however, has been variable and a broad range of sensitivities and specificities have been reported. The purpose of this investigation was to evaluate the usefulness of (111)In-labelled leucocytes scintigraphy in the detection of a cause of FUO in the light of a changing spectrum of diseases causing it and advances in investigational techniques. MATERIALS AND METHODS: Sixty-one patients with a clinical diagnosis of FUO underwent whole-body (111)In-troponolate-labelled leucocyte scintigraphy in our department over a 2-year period between February 2004 and February 2006. Of these, 54 patients were retrospectively reviewed to identify a cause of FUO. Other parameters such as C-reactive protein (CRP), leucocyte count and radiological findings were also evaluated. RESULTS: Leucocyte scintigraphy was found to be true positive in 12 patients, true negative in 24 patients, false positive in 10 patients and false negative in eight patients. The overall sensitivity of scintigraphy was 60%, specificity 71%, positive predictive value 55%, and negative predictive value 75%. There was no difference in the scintigraphic sensitivity between patients with spontaneous FUO and those with post-operative FUO although the latter showed a higher specificity and PPV. CRP and leucocyte count did not differ significantly between patients with true positive and true negative scintigrams. Overall, 83% of patients with abnormal radiological examinations had positive findings on scintigraphy and 87% of patients with negative findings on radiology had normal scintigraphy. CONCLUSION: Despite changes in disease spectrum and advances in investigational techniques, our results suggest that (111)In-leucocyte scintigraphy is still a useful technique in establishing the cause of FUO. A higher PPV of this test in post-operative situations makes it especially applicable in this category of patients. Equally, the higher NPV in patients with spontaneous FUO virtually excludes infection/inflammation. Finally, a higher pre-test probability based on the radiological tests seems to be important in the optimal use of leucocyte imaging.

Dual-isotope lymphoscintigraphy using albumin nanocolloid differentially labeled with 111In and 99mTc.

The aim of this study was to develop and evaluate 111In- and 99mTc-labeled derivatives of albumin nanocolloid (NC) for dual-label lymphoscintigraphy to allow simultaneous comparison of lymphatic flow from different tissue planes draining a tumour bed for accurate identification of sentinel lymph nodes (SLN). Using the chelator, p-isothiocyanatobenzyl-1,4,7, 10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 111In-DOTA-NC and 99mTc-DOTA-NC were compared in vitro with respect to stability of labeling, colloidal status and particle size, then in vivo by measuring their clearance rates from a subcutaneous injection depot. 111In-DOTA-NC and 99mTc-DOTA-NC were indistinguishable on the basis of in vitro criteria. Their in vivo clearance rates, however, were disparate (0.0015 to 0.075 min(-1) for 111In and 0.0072 to 0.067 min(-1) for 99mTc), 111In being faster in three studies and markedly slower in three. This demonstrates that even when dual-labeled radiotracers behave identically in vitro, they will not necessarily do so in vivo. Further work is needed to develop dual-labeled NC.

Imaging of lymphatic vessels in breast cancer-related lymphedema: intradermal versus subcutaneous injection of 99mTc-immunoglobulin.

OBJECTIVE: The disordered physiology that results from axillary lymph node clearance surgery for breast cancer and that leads to breast cancer-related lymphedema is poorly understood. Rerouting of lymph around the axilla or through new pathways in the axilla may protect women from breast cancer-related lymphedema. The aim of the study was to compare intradermal with subcutaneous injection of technetium-99m ((99m)Tc)-labeled human polyclonal IgG (HIG) with respect to lymphatic vessel imaging. MATERIALS AND METHODS: Six women with breast cancer-related lymphedema underwent unilateral upper limb lymphoscintigraphy, using a web space injection of (99m)Tc-labeled HIG, after intradermal and subcutaneous injections on separate occasions. Multiple sequential images were obtained of the affected upper limb and torso over 3 hr on each occasion. Accumulation of activity in blood was quantified from venous blood samples taken from the opposite arm. RESULTS: Imaging after intradermal injection clearly showed discrete lymphatic vessels in five of six patients, in contrast to imaging after subcutaneous injection, which did not show any discrete vessels in any patient. Intradermal injection resulted in more rapid visualization of cutaneous lymph rerouting than subcutaneous injection in six of six patients. Recovery of injected (99m)Tc-labeled HIG in venous blood was greater after intradermal injection in six of six patients. CONCLUSION: In patients with breast cancer-related lymphedema, lymphatic vessels are more clearly depicted after intradermal than subcutaneous injection as a result of direct access of radiotracer to dermal lymphatics. This finding has implications for imaging lymphatic vessel regeneration and lymph rerouting.

Short-term effects of axillary lymph node clearance surgery on lymphatic physiology of the arm in breast cancer.

It is not known why some women develop breast cancer-related lymphedema (BCRL) of the arm, whereas others having similar treatment do not. We speculated that increased uptake of protein into local blood may protect against BCRL. Sixteen women were given bilateral subcutaneous hand webspace injections of polyclonal immunoglobulin (HIgG), (99m)Tc-HIgG on one side and (111)In-HIgG on the other, before and 3 mo after axillary clearance surgery. The rates of clearance of activity from the depot (k) and accumulation in central blood (b(contra)) were measured using a scintillation probe and bilateral antecubital vein blood sampling, respectively. Activity accumulating in blood ipsilateral to the injected side, in excess of central blood activity (b(ipsi)) was also calculated as a measure of local vascular uptake. The k correlated with b(contra), but neither changed in response to surgery. However, b(ipsi) for injections of (99m)Tc-HIgG into the affected arm increased in all seven patients in whom data were available (0.018 +/- 0.006 to 0.038 +/- 0.007%/min; P < 0.05); indeed, in five of these seven, b(ipsi) paradoxically exceeded b(contra), and none developed BCRL at 3-yr follow-up. We conclude that uptake of protein into local blood and/or proteolysis increases after axillary surgery and may protect against BCRL.

Quantification of disease activity in patients undergoing leucocyte scintigraphy for suspected inflammatory bowel disease.

PURPOSE: Whole-body gamma camera counting is an alternative to faecal 111In collection for quantifying disease activity in inflammatory bowel disease (IBD) but requires administration of imaging activities of 111In. The aim of this study was to explore a dedicated whole-body counter which requires 20-fold less activity than gamma camera counting. METHODS: Thirty patients with known or suspected IBD received 99mTc-granulocytes (approximately 200 MBq) and 111In-granulocytes (approximately 0.5 MBq). The 99mTc-cells were injected 45 min after the 111In-cells and immediately after a baseline 111In whole-body count. The decay-corrected count at 120 h was expressed as a fraction of baseline to give whole-body 111In retention (WBR). One patient was excluded as the injected cells were non-viable. RESULTS: Median 45-min intravascular 111In recovery was 35% in patients compared with 43% in six normal volunteers (p<0.05). WBR in normals ranged from 91% to 93% and in 11 patients with negative 99mTc imaging from 87% to 96%. Only one of 11 patients with negative imaging had WBR <90% while 11/12 patients with WBR <90% had abnormal imaging. Ten of 13 patients with Crohn's disease (CD) had abnormal imaging. The lowest WBR in these ten was 77% and six had values of >90%. There was a significant association between WBR and 99mTc image grade (Rs=0.73, p<0.002) in 16 patients without CD, but not in 13 patients with CD (Rs=0.54, p>0.05). CONCLUSION: Dedicated whole-body counting is able to quantify disease activity in IBD but may give normal values in active CD.

Measurement of the extraction fractions of nanocolloid and polyclonal immunoglobulin by axillary lymph nodes in patients with breast cancer.

BACKGROUND: 99mTc nanocolloid (99mTc-NC) is the most widely used tracer for lymphoscintigraphy, although others have been proposed, including radiolabelled proteins such as human serum albumin and polyclonal human immunoglobulin G (HIG). The extraction fraction of such tracers by individual nodes is clearly important but has not previously been measured in humans. METHODS: Patients scheduled for axillary clearance surgery (three groups) received dual-labelled radiotracers 2-4 h before surgery: group 1 (3 patients) received 99mTc-NC (10 MBq) and 111In-HIG (2 MBq) as a mixture (0.2 ml) into the breast parenchyma above the primary tumour; group 2 (3 patients) received 99mTc-HIG (10 MBq) and 111In-HIG (2 MBq) as a mixture (0.2 ml) into the breast parenchyma above the primary tumour; and group 3 (4 patients) received 99mTc-HIG (10 MBq) and 111In-HIG (2 MBq) separately (both 0.2 ml) into the breast parenchyma above the tumour and the intradermal plane at the areola. All resected nodes were counted for Tc and In in a well-type scintillation counter. In group 1, nodes were ranked according to their Tc uptake. In groups 2 and 3, nodes were ranked separately according to their respective Tc and In uptakes. If nodes are arranged in linear order and each node extracts a constant fraction of incoming tracer, then the activity in the nodes would decrease exponentially with an individual nodal extraction fraction, E, equal to 1-e(-k), where k is the rate constant of decrease. RESULTS: In the first group, 99mTc-NC and 111In-HIG identified the same sentinel and second echelon nodes. The observed decrease in nodal activity was exponential in all groups, at least for the first five nodes. Average values for E, based on the first five nodes were 0.69 (range 0.57-0.89; n=3) for 99mTc-NC and 0.45 (0.15-0.70; n=17) for HIG (irrespective of label) (Wilcoxon rank sum, P=0.02). With respect to HIG, there was no significant difference in E between 99mTc and 111In or between deep and superficial injections in group 3. CONCLUSION: Although HIG has an extraction fraction less than 99mTc-NC, the value of E is still high enough to make HIG a useful tracer for lymphoscintigraphy, especially for identifying second echelon nodes in addition to sentinel nodes and for imaging lymphatic vessels as well as lymph nodes.

Local vascular access of radioprotein injected subcutaneously in healthy subjects and patients with breast cancer-related lymphedema.

UNLABELLED: The aim of the study was to use dual-isotope lymphoscintigraphy in healthy volunteers and women with breast cancer-related lymphedema (BCRL) to detect and quantify transport of radiolabeled protein from a subcutaneous injection depot to local blood vessels as a potential mechanism of protection against edema resulting from treatment to the axilla. METHODS: A total of 29 subjects and 18 women with a history of BCRL received bilateral subcutaneous injections of human IgG (HIgG) in the second dorsal web space of each hand, (99m)Tc-HIgG on one side and (111)In-HIgG on the other. In 8 further healthy subjects, epinephrine was administered with the labeled HIgG. Radioactivity at each depot was measured at regular intervals for a total of 3 h using a collimated sodium iodide scintillation detector, and radioactivity in venous blood sampled from both arms was measured using an automatic sample counter. Ipsilateral blood time-concentration curves were corrected for recirculating activity by subtraction of the simultaneous contralateral concentration, to define the component of ipsilateral blood resulting from local vascular access of radioprotein. Accumulation of activity in blood was expressed in relation to injected activity and activity that had left the depot and was calculated as a function of time-in systemic blood, by multiplying contralateral concentrations by an estimate of the subject's blood volume, and in ipsilateral blood, by using indicator dilution theory and an assumed forearm blood flow of 20 mL/min. RESULTS: (99m)Tc-HIgG and (111)In-HIgG behave almost identically with respect to depot clearance and accumulation in contralateral venous blood, with or without epinephrine, which reduced both depot clearance and blood accumulation rate. Moreover, a side-to-side correlation with respect to contralateral accumulation was present in healthy subjects, was not abolished by epinephrine, and was maintained in the face of asymmetric accumulation in BCRL. Contralateral accumulation of radioprotein was reduced in BCRL after injection into the affected side only when the hand was involved. In contrast to contralateral sampling, ipsilateral time-concentration and accumulation profiles were consistent with instability of (111)In-HIgG and rapid local vascular access of small amounts of protein-free (111)In. Experiments based on precipitation of protein with trichloroacetic acid confirmed relatively high levels of unbound ipsilateral (111)In, especially in samples obtained early after injection. Substantial accumulation of protein-bound (99m)Tc was observed in ipsilateral blood, with a time course similar to that of contralateral accumulation. Positive correlation between ipsilateral and contralateral blood (99m)Tc activity was observed at all time points, often significantly, in contrast to (111)In, for which it was negative at all time points. Ipsilateral accumulation of (99m)Tc adjusted for activity that had left the depot was unchanged with respect to the affected arm in BCRL patients. CONCLUSION: Whereas (111)In-HIgG and (99m)Tc-HigG are interchangeable for measurement of depot clearance and contralateral venous accumulation rates, ipsilateral sampling is much more sensitive to protein-free radionuclide and detects significant differences resulting from some instability of (111)In-HIgG. On the basis of (99m)Tc data, there appears to be substantial local vascular access of radioprotein within the arm, both in healthy subjects and in patients with BCRL, through either lymphaticovenous communications or direct transendothelial transport.

Finding an optimal method for imaging lymphatic vessels of the upper limb.

Lymphoscintigraphy involves interstitial injection of radiolabelled particulate materials or radioproteins. Although several variations in the technique have been described, their place in clinical practice remains controversial. Traditional diagnostic criteria are based primarily on lymph node appearances but in situations such as breast cancer, where lymph nodes may have been excised, these criteria are of limited use. In these circumstances, lymphatic vessel morphology takes on greater importance as a clinical endpoint, so a method that gives good definition of lymphatic vessels would be useful. In patients with breast cancer, for example, such a method, used before and after lymph node resection, may assist in predicting the development of breast cancer-related lymphoedema. The aim of this study was to optimise a method for the visualisation of lymphatic vessels. Subcutaneous (sc) and intradermal (id) injection sites were compared, and technetium-99m nanocolloid, a particulate material, was compared with (99m)Tc-human immunoglobulin (HIG), which is a soluble macromolecule. Twelve normal volunteers were each studied on two occasions. In three subjects, id (99m)Tc-HIG was compared with sc (99m)Tc-HIG, in three id (99m)Tc-nanocolloid was compared with sc (99m)Tc-nanocolloid, in three id (99m)Tc-HIG was compared with id (99m)Tc-nanocolloid and in three sc (99m)Tc-HIG was compared with sc (99m)Tc-nanocolloid. Endpoints were quality of lymphatic vessel definition, the time after injection at which vessels were most clearly visualised, the rate constant of depot disappearance ( k) and the systemic blood accumulation rate as measured by gamma camera imaging over the liver or cardiac blood pool. Excellent definition of lymphatic vessels was obtained following id injection of either radiopharmaceutical, an injection route that was clearly superior to sc. Differences between radiopharmaceuticals were less clear, although after id injection, (99m)Tc-HIG gave images that were marginally but significantly better than those given by (99m)Tc-nanocolloid. Image quality correlated inversely with time after injection at which the best image was obtained, consistent with the notion that good vessel definition was dependent on a "narrow" bolus width. k was approximately three times higher after id injection than after sc injection but it was not significantly different between radiopharmaceuticals for either injection route. Intradermal (99m)Tc-HIG gave a cardiac blood pool signal that, over the first 60 min, increased about five times faster than that with sc (99m)Tc-HIG, but no clear difference was observed in the rate of increase in hepatic activity between id (99m)Tc-nanocolloid and sc (99m)Tc-nanocolloid. We conclude that id injection provides rapid access of radiotracers to lymphatic vessels, which is ideal for imaging lymphatic vessel morphology. (99m)Tc-HIG is marginally superior to nanocolloid for this purpose and, in drainage basins from which lymph nodes have been excised, is not handicapped by a potentially inferior ability, compared with radiocolloid, to image lymph nodes.