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Drainage of lymph from the foreleg to the superficial cervical lymph node in sheep.

The superficial cervical (prescapular) node in sheep is large and readily accessible. It lies medial to the omotransverse muscle and 40 to 60 mm cranial to the ventral end of the scapular spine. Lymph vessels reaching this node from the foreleg all enter its ventral half. These lymphatics can be approached surgically opposite the elbow joint where they lie adjacent to the cephalic vein. Carbon particles infused into a single afferent lymphatic were restricted to a small segment of the ventrocranial quadrant of the node. When Evans' Blue dye, or a suspension of carbon particles, was injected subcutaneously just above the hoof they were carried to the node in two to five afferent lymphatics and were found mainly in the ventrocranial quadrant. A knowledge of the anatomy of the superficial cervical node and its afferent lymphatics may be of value in immunological studies, especially where it is desired to introduce antigen into a known part of a node and to leave the remainder for control observations.

Animals↗

Sonography of neck lymph nodes. Part II: abnormal lymph nodes.

Assessment of cervical lymph nodes is essential for patients with head and neck carcinomas, and ultrasound is a useful imaging technique. Sonographic features that help distinguish between the causes of neck lymphadenopathy, including grey scale and Doppler features, are discussed. In addition to the distribution and location of nodes, the useful grey-scale features are: size, shape, internal architecture, intranodal necrosis, absence of hilar structure and calcification. The useful Doppler features are: distribution of vascularity and intranodal resistance. Ancillary features such as oedema of soft tissue and nodal matting are particularly helpful to identify tuberculous nodes.

Calcinosis↗

Application of the popliteal lymph node assay in immunotoxicity testing: complementation of the direct popliteal lymph node assay with flow cytometric analyses.

The popliteal lymph node assay (PLNA) has been proposed to measure the immunosensitizing potential of chemicals. The direct PLNA detects an immunomodulating effect but does not give insight into the mode of action of the chemical under test. Modifications of this test have been proposed, but they are difficult to perform in routine toxicity testing and require many animals. In the present investigation the direct PLNA was extended with the flow cytometric determinations of: (a) lymphoblasts; (b) the phenotyping of lymphoid subpopulations; (c) the determination of expression of proliferation/activation markers CD25, CD69 and CD62L/CD44 and (d) the analysis of intracellular cytokines interferon gamma, interleukin 2 and interleukin 4. Streptozotocin, hydrazine, HgCl2 and trinitrobenzene sulfonic acid were used as model chemicals. The different mode of action of these substances was well documented by the techniques applied. As the proposed flow cytometric methods can easily be performed and do not require additional test animals this complementation of the direct PLNA seems a promising approach in immunotoxicity testing.

Allergens↗

Tumour thickness as a predictor of occult lymph node metastases in patients with stage I and II melanoma undergoing sentinel lymph node biopsy.

BACKGROUND: Sentinel lymph node biopsy (SLNB) is a minimally invasive procedure used accurately to stage nodal basins at risk of occult metastases. There are no data as yet to show a survival benefit from SLNB and its use remains controversial. If Breslow thickness of the tumour correlates well with positive SLNB, it could be used to select patients for SLNB. METHODS: A quantitative systematic review of published studies on SLNB in patients with melanoma available by September 2001 was performed. RESULTS: Twelve studies containing 4218 patients with stage I and II melanoma were identified; 17.8 (95 per cent confidence interval 16.7 to 19.0) per cent of patients had nodal micrometastases detected by SLNB. The incidence of micrometastasis in sentinel nodes correlated directly with Breslow tumour thickness; it was 1.0 per cent for lesions of less than or equal to 0.75 mm, 8.3 per cent for 0.76-1.50 mm, 22.7 per cent for 1.51-4.0 mm and 35.5 per cent for more than 4.0 mm. CONCLUSION: The Breslow thickness of primary melanoma predicts the presence of a sentinel node metastasis. The published data are not sufficient to demonstrate a correlation between other known prognostic indicators and a positive SLNB.

Humans↗

The in vitro differentiation of mast cells. Cultures of cells from immunized mouse lymph nodes and thoracic duct lymph on fibroblast monolayers.

When cells from lymph nodes or thoracic duct of mice hyperimmunized with protein antigens are cultivated on embryo monolayers in the presence of the antigen, numerous clones of mast cells appear. The histochemical and ultrastructural characteristics of the cells permit their identification as mast cells and distinguish them from the phagocytic histiocytes that usually arise in abundance in similar cultures from unimmunized mouse cells or from immunized mouse cells cultured in the absence of the antigen. Only a few colonies of mast cells appeared in the latter cultures. The basis for the induction of mast cell differentiation is not known.

Animals↗

Selective disruption of lymphotoxin ligands reveals a novel set of mucosal lymph nodes and unique effects on lymph node cellular organization.

Lymphotoxin (LT) provides a critical signal for the genesis of lymph nodes (LN) in mice. Here we show that mice treated in utero with LT beta-R-Ig, which binds to the membrane LT alpha 1 beta 2 heterotrimer, lacked most LN, yet retained a set of mucosal surface draining LN. Since mice genetically deficient in LT alpha lack all LN, including the mucosal set, we hypothesize that a novel LT alpha-dependent pathway controls their genesis. This novel set of mucosal LN cannot be discriminated on the basis of addressin expression. The discovery of LN in mice treated with LT beta-R-Ig fusion protein in utero allowed us to compare the roles of membrane LT alpha beta or soluble LT alpha/tumor necrosis factor (TNF) in the development of cellular organization in LN and spleen. Our results indicate that both membrane LT alpha beta and soluble LT alpha/TNF mediate T-B cell segregation and the organization of B cell follicles in spleen and LN. Interestingly, while antagonism of membrane LT alpha beta or soluble LT alpha/TNF prevented germinal center (GC) formation in spleen, antagonism of soluble LT alpha/TNF had no effect on LN formation. The data suggest that multiple LT/TNF ligands control B cell follicle organization in the spleen and LN of adult mice, and that the requirements for LT/TNF ligands in GC formation are distinct in the different lymphoid organs.

Animals↗

The architecture of rat lymph nodes. IV. Distribution of ferritin and colloidal carbon in the draining lymph nodes after foot-pad injection.

Rat popliteal and the iliac lymph nodes were examined after foot-pad injections of colloidal carbon and ferritin-tetramethylrhodamine isothiocyanate. Carbon rapidly entered medullary sinuses, but the entry of carbon into the interstitium was prohibited by avid phagocytosis, by sinus macrophages and by the lymphoendothelium, which apparently formed a barrier to diffusion. In contrast, little carbon was phagocytosed in subcapsular sinuses, from where the particles entered the underlying cortex through holes in the lymphoendothelium created by penetrating frilly cells. The distribution of ferritin was similar to that of carbon. Both carbon and ferritin localized poorly in follicles; however, preinjection of specific antibody caused enhanced follicular localization of ferritin. By electron microscopy clusters of ferritin molecules were found on the surface of dendritic cells. These cells showed different morphology from that of the interdigitating cells of the paracortex. The latter cells did not bind ferritin to their surface, even in the presence of specific antibody.

Animals↗

The sentinel lymph node concept in prostate cancer - first results of gamma probe-guided sentinel lymph node identification.

OBJECTIVE: The goal of this study was to show lymphatic drainage and to verify the validity of lymphoscintigraphy for the identification of the sentinel lymph node (SLN) in prostate cancer. Furthermore, the question is to be raised whether the standardized pelvic lymphadenectomy is a sufficient means for also detecting solitary micrometastases. PATIENTS AND METHODS: Eleven patients with prostate cancer received a sonographically controlled, transrectal administration of a technetium-99m colloid injected directly into the prostate 1 day prior to pelvic lymphadenectomy. 20 min later the dynamic lymphoscintigraphy was carried out. During surgery, the SLNs were identified by using a gamma probe. The standard pelvic lymphadenectomy was performed after removal of the SLN. RESULTS: In 3 of 4 patients with micrometastasis the spread of the tumor could exclusively be found in those nodes which had been identified as SLNs by means of scintigraphy by combining preoperative lymphoscintigraphy and intraoperative gamma probe detection. In 2 cases, the pathologically proved SLNs were situated at the anteromedial region of the internal iliac artery, thus being located outside of the standard pelvic lymphadenectomy area. In 1 patient, however, the micrometastasis was found beyond those nodes which had been identified as SLN intraoperatively. CONCLUSIONS: In the future, we expect the restriction of pelvic staging lymphadenectomy to scintigraphically proved SLN. The perioperative morbidity may be reduced by increasing the sensitivity of the detection of micrometastases. Our data confirm earlier perceptions, according to which even modified standardized pelvic lymphadenectomy is considered insufficient in terms of the detection of micrometastases.

Aged↗

Lymph node staging in non-small-cell lung cancer with FDG-PET scan: a prospective study on 690 lymph node stations from 68 patients.

PURPOSE: To compare the accuracy of computed tomography-(CT) scan and the radiolabeled glucose analog 18F-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) visually correlated with CT (PET + CT) in the locoregional lymph node (LN) staging of non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Sixty-eight patients with potentially operable NSCLC underwent thoracic CT, PET, and invasive surgical staging (ISS). Imaging studies were read prospectively and blinded to the surgical and pathologic data. A five-point visual scale was used for the interpretation of LNs on PET. Afterwards, with knowledge of the pathology, the relationship between standardized uptake values (SUVs) and the presence of metastasis in LNs was explored in a receiver operating characteristic (ROC) analysis, and the likelihood ratios (LRs) for SUVs of LNs were determined. RESULTS: ISS was available for 690 LN stations. CT correctly identified the nodal stage in 40 of 68 patients (59%), with understaging in 12 patients and overstaging in 16 patients. PET + CT was accurate in 59 patients (87%), with understaging in five patients and overstaging in four patients. In the detection of locally advanced disease (N2/N3), the sensitivity, specificity, and accuracy of CT were 75%, 63%, and 68%, respectively. For PET + CT, this was 93%, 95%, and 94% (P = .0004). In the ROC curve, the best SUV threshold to distinguish benign from malignant LNs was 4.40. The analysis with this SUV threshold was not superior to the use of a five-point visual scale. The LR of a SUV less than 3.5 in an LN was 0.152; for a SUV between 3.5 and 4.5, it was 3.157; and for a SUV greater than 4.5, it was 253.096. CONCLUSION: PET + CT is significantly more accurate than CT alone in LN staging of NSCLC. A five-point visual scale is as accurate as the use of an SUV threshold for LNs in the distinction between benign and malignant nodes. The very high negative predictive value of mediastinal PET could reduce the need for mediastinal ISS in NSCLC substantially.

Adult↗

Factors predictive of tumor-positive nonsentinel lymph nodes after tumor-positive sentinel lymph node dissection for melanoma.

PURPOSE: Approximately 20% of sentinel node (SN) positive melanoma patients have additional non-SN (NSN) metastasis. The rationale for this study was to identify the factors associated with additional nodal disease, as a method to determine which patients may most benefit from completion lymph node dissection (CLND). PATIENTS AND METHODS: During 1990 to 2002, 1,599 patients have undergone SN biopsy at our institute. 19.5% underwent CLND for tumor-positive SN. One hundred ninety-one of these patients had clinicopathologic information available for review. Univariate analyses used chi2 test, Wilcoxson rank sum test, and chi2 test for trend. Multivariate analyses used logistic regression and Wald test. RESULTS: Forty-six (24%) patients had tumor-positive NSN. Univariate analyses showed that primary thickness (Breslow and Clark), primary site, SN tumor size, and number of tumor-positive SNs were significantly associated with tumor-positive NSN. Multivariate analysis (167 patients), confirmed that Breslow and SN tumor size were independently predictive. Sex, histology, ulceration, mitotic index, and SN basin location were not predictive. Risk stratification by the number of prognostic factors present (Breslow > or = 3 mm and SN tumor size > or = 2 mm) showed that probability of finding tumor-positive NSN was 12.3% in the low-risk group (0 factors), 30.9% in the intermediate-risk group (1 factor), and 41.9% in the high-risk group (2 factors). CONCLUSION: Thicker primary and larger SN tumor size are factors that correlate best with tumor-positive NSN. Although none of these factors are absolutely predictive of residual nodal disease, these factors must be strongly considered if the SN contains metastasis, as they provide enhanced risk assessment for NSN tumor-positivity.

Adolescent↗

[Regional neck lymph nodes and primary tumors. 1. Neck lymph node metastases].

The metastasis are situated in the primary regional lymphnodes. Preoperative palpation of the neck gives unreliable results. The extent of the primary tumor has no correlation to the occurence of metastasis. When the tumor is localized in the supraglottic, transglottic or in the tongue base region the frequency of metastasis in lymph nodes is approximately equal; more than half of them have metastasis. When the tumor is localised in sinus piriformis or in the marginal region a still higher frequency of metastasis is found.

Head and Neck Neoplasms↗

Stimulation of cells from a non-invaded and an invaded lymph node with a HER-2+ tumor with peptides corresponding to T-cell epitopes E75 and G89 induced expansion of central memory cells (TCM) from the metastasis-negative lymph nodes.

Breast cancer metastasizes from the primary site to the axillary lymph nodes (LN). It is unknown whether tumor metastasis abolishes or enhances the ability of LN cells to develop a specific response to the Ag expressed by the tumor, and whether an immune response to the same Ag is present in the tumor-free LN. We stimulated lymphocytes from a metastasis negative (Met-) and a metastasis positive (Met+) LN, invaded by a HER-2+ tumor, from the same patient, with HER-2 peptides E75 (369-377) and G89 (776-778). E75 define a CTL epitope presented by HLA-A2, while G89 define a CD4+ cell recognized epitope. Met- LN responded to E75+G89 with higher expansion of E75 TCR+ CD45RO+ CCR7- (CCR7-) and E75-TCR+ CD45RO+ CCR7+ (CCR7+) cells than Met+ LN. Stimulation with E75+G89 induced a significant increase in CCR7+ cells in Met- LN compared with Met+ LN. The levels of IFN-alpha and IL-15 were higher in Met- LN cultures stimulated with E75+G89 than in Met+ LN cultures. This increase did not correlate with the levels of induction of IFN-gamma, IL-4, and IL-10. The finding of higher expansion of Ag specific CCR7+ cells and of differentiation to CCR7- cells, which define the TCM and TEM subsets respectively, in Met- LN, by G89 is novel for tumor systems. This may have implications for preventative vaccination strategies for breast and ovarian cancer.

Breast Neoplasms↗

Lymphocyte recognition of lymph node high endothelium: adhesive interactions determining entry into lymph nodes.

Our interpretation of these results is that lymphocyte surface HEBF is composed of high endothelial adhesion molecules which mediate cell-cell interactions that result in lymphocyte entry from blood into lymph nodes. The evidence that anti-HEBF antibody does not interfere with lymphocyte entry into PP is intriguing, since migration into this tissue as well as LN, occurs via HEV. This suggests that high endothelial cells differ in LN and PP at least with respect to the specificity of surface molecules involved in lymphocyte adherence. It could be that separate lymphocyte subpopulations express receptors for these two types of high endothelium. If this explanation is correct, then cells negatively selected using this antibody should be capable of binding to HEV of PP but not HEV of LN. However, if receptors for both high endothelial types are present on the same lymphocyte, then it is unlikely that such selection would yield cells exhibiting tissue specificity. Our observations are consistent with previous findings which suggested that high endothelial cells of mouse LN and PP exhibit differences in lymphocyte binding properties. For example, it has been reported that certain murine lymphomas bind to HEV of either peripheral LN or PP and that, to a limited degree, this preference is a property of most B and T cells [Butcher et al, 1980; Stevens et al, 1982]. Thymus and bone marrow cells show only low levels of HEV binding and less than 10% of such cells react with anti-HEBF antibody. During differentiation lymphocytes appear to acquire surface components which mediate high endothelial adhesion and this event is associated with the appearance of surface molecules recognized by anti-HEBF antibody. This suggests that it is the expression of these surface molecules during differentiation which confers high endothelial recognition properties on lymphocytes and that this mechanism plays a role in adhesive interactions leading to entry of both T and B cells into LN.

Animals↗

The relationship between tissue fluid and lymph. II. Enzymes in tissue fluid and lymph.

In rabbits both under normal conditions and after burning the activity of intracellular enzymes (LDH, GOT) is higher in tissue fluid than in the peripheral (leg) lymph. The relationship between tissue fluid and lymphatic enzyme concentrations is non-linear. The observations suggest the presence of at least two distinct compartments in tissue fluid. The first compartment forms the pathway taken by the fluid and protein leaving the blood capillaries and is directly drained by the lymphatics. The second compartment is the 'true' tissue fluid, contains the extravascular protein pool and it is in dynamic equilibrium with the first compartment.

Animals↗

Dramatic hyperplasia of mtv-2+ lymph node grafts in mtv-2- recipients and selective stimulation of V beta 14+ T cells in recipients' lymph nodes in the DDD mouse.

DDD/1 (DDD) mice contrast strikingly with DDD-mtv-2/mtv-2 (DDD-mtv-2) congenics in their marked lymph node (LN) T cell paucity. To clarify the possible difference in LN function between them, reciprocal LN grafting experiments were conducted. DDD-mtv-2 LN grafts in DDD recipients underwent hyperplasia as dramatic as 10-to 20-fold increase in weight between 3 and 4 wk after implantation. Lymphoid cells in hyperplastic LN grafts were of recipient origin. Similar hyperplasia of mtv-2-heterozygous LN grafts also occurred on various hybrid backgrounds involving DDD mice. Moreover, LN grafts from BALB/c mice infected with mtv-2-derived exogenous mouse mammary tumor virus (MMTV) swelled in MMTV-free BALB/c recipients. Genetic analysis of DDD x (DDD x DDD-mtv-2)F1 backcross progeny demonstrated that LN hyperplasia was closely linked to mtv-2. The frequencies of V beta 5+ and V beta 8+ T cells unresponsive to mtv-2-encoded superantigen (SAg) changed with practically the same kinetics in both LN grafts and recipients' LN. Thus, the cells responsible for LN hyperplasia were polyclonal. V beta 14+ CD4+ cells responsive to mtv-2 SAg were specifically stimulated in recipients' LN but selectively deleted in hyperplastic LN grafts. DDD mice carrying hyperplastic mtv-2+ LN grafts or pretreated with mtv-2+ spleen cells developed an unresponsive state in terms of influx of mtv-2- lymphoid cells into mtv-2+ LN grafts. These results indicate that mtv-2 gene products including SAg may stimulate mtv-2- lymphoid cells of recipients and cause them to migrate into mtv-2+ LN grafts in a nonspecific manner with resulting LN hyperplasia.

Animals↗

Mucosal or targeted lymph node immunization of macaques with a particulate SIVp27 protein elicits virus-specific CTL in the genito-rectal mucosa and draining lymph nodes.

The major routes of HIV transmission are through the rectal and cervico-vaginal mucosa. To prevent dissemination of HIV to the regional lymph nodes (LNs), an effective vaccine may need to stimulate CTL in the rectal or genital tract and the draining LNs. We report that mucosal immunization by the recto-oral and vagino-oral route or s.c. immunization targeting the iliac LNs with a particulate SIVp27:Ty-VLP vaccine elicits SIVgag-specific CTL in the regional LNs as well as in the spleen and PBMC. Targeted LN immunization with this vaccine elicited MHC class I-restricted CD8+ CTL responses, and the highest frequency of CTL was found in the iliac LNs. Moreover, SIVgag-specific CTL activity was detected in short term T cell lines established in mononuclear cells eluted from the rectal and cervico-vaginal mucosa. The relative frequency of CTL in short term cell lines prepared from the rectal mucosa (21/113 or 18.6%) was similar to that obtained from the cervico-vaginal mucosa (16/79 or 20.3%). Examination of the relative frequency of CTL to the T cell epitopes residing within SIVp27 showed a higher frequency in iliac LN cells to peptide aa 41-70 than in that to peptide aa 121-150, and this was significant after both recto-oral (chi-squared 6.500, p < 0.02) and vagino-oral (chi-squared = 10.391, p < 0.01) immunization. In contrast, the relative frequency of CTL in PBMC to peptide aa 41-70 (15.5%) was comparable to that elicited by peptide aa 121-150 (17.6%). This study provides novel evidence that mucosal or targeted LN immunization can generate anti-SIV CTL in the rectal and genital mucosa, in the draining LNs, and in the central lymphoid system.

Animals↗