Cannulated Lymph Vessels Research Articles

Large increase of Langerhans cells in human skin lymph derived from irritant contact dermatitis.

In order to monitor the kinetics of Langerhans cells in the afferent lymph during contact dermatitis, a superficial peripheral lymph vessel draining the skin of the upper and medial part of the foot was cannulated by means of microsurgery on the lower leg of four healthy volunteers. After 2 days an irritant contact dermatitis was induced by application of 10% sodium lauryl sulphate to the area of skin drained by the cannulated lymph vessel. Three days later the spontaneously regressing skin reaction was treated with clobetasol propionate in two of the subjects. Lymph was collected twice daily for 8 days. Langerhans cells were identified by immunofluorescence microscopy of cytocentrifuge slide preparations from the lymph, using a monoclonal anti-CD1a antibody. In the late phase of the contact dermatitis the output, i.e. both the absolute number and the percentage of Langerhans cells in the lymph dramatically increased. At the end of the experiment, when there were no remaining clinical signs of contact dermatitis, the Langerhans cell output still markedly exceeded the initial values. These results are the first direct evidence in humans that migration of Langerhans cells from the skin to the regional lymph nodes is a major feature of irritant contact dermatitis.

Immune events in skin. I. Spontaneous cluster formation of dendritic (veiled) cells and lymphocytes from skin lymph.

To investigate the mechanism of spontaneous attachment of afferent lymph lymphocytes to dendritic cells, cells from canine skin lymph were used. There were 3.3 +/- 2.8% of veiled cells in clusters found in lymph flowing from the cannulated lymph vessel. The number of clusters forming ex vivo in the collected lymph samples increased as a function of time and was temperature dependent. Incubation of cells with proteolytic enzymes or monosaccharides did not alter cell interactions. The ability of veiled cells to bind lymphocytes was independent of divalent cations but reduced by xylocaine and retinoic acid. Among steroids only methylprednisolone showed an inhibitory effect on cluster formation. Indomethacin and acetylsalicylic acid had no blocking activity on cell binding. Also, no effect was seen after treatment with cyclosporine A and azathioprine. An enhanced cluster formation after desialation with neuraminidase was observed. The desialated cells were cultured in order to study their stimulatory and accessory cell functions. No enhancement of autologous mixed leucocyte reaction was seen, but a significantly higher responsiveness to a suboptimal dose of phytohaemagglutinin was observed. The N-ase-mediated non-specific cell attachment could be abrogated by cell washing or treatment with EDTA or xylocaine. This study indicates that cluster formation by skin lymph veiled cells and lymphocytes is a spontaneous process which cannot be controlled by means usually effective in regulating the in vitro induced clustering of antigen-stimulated cells.

Isolation of human skin-derived lymph: flow and output of cells following sodium lauryl sulphate-induced contact dermatitis.

By means of microsurgery a peripheral subcutaneous lymph vessel draining a defined skin area was isolated and cannulated on the lower leg of six healthy volunteers. Lymph was collected over a period of 8 days. During the first 2 days baseline values for lymph flow and output of cells were established. A contact dermatitis was then induced in the drained skin area by the application of 10% sodium lauryl sulphate. All six probands developed a mild to moderate irritant contact dermatitis. Lymph flow as well as output of cells increased with the intensity of the skin reaction. Subsequent local treatment with clobetasol propionate decreased the cell output, but the lymph flow increased further. Neither lymph flow nor output of cells returned to the initial baseline values at the end of the study, when the clinical signs of contact dermatitis had completely disappeared. During the experiment significant individual variations were found, with means ranging from 0.10 to 0.48 ml/h for lymph flow and from 8700 to 174000/h for cells, which probably depended mainly on the different topographies and calibres of the cannulated lymph vessels.

Lymph flow from edematous dog lungs.

We measured the flow rate (QLV) from cannulated lung lymph vessels in anesthetized dogs. Low-resistance lymph cannulas were used and the vessels were cannulated at the lung hilus. When we increased left atrial pressure to 42.9 +/- 5.7 (SD) cmH2O (base line = 6.6 +/- 4.6 cmH2O), the lungs became edematous and QLV increased from a base line of 20.4 +/- 21.5 microliters/min to 388 +/- 185 microliters/min. QLV plateaued at the higher level. We also measured the relationship between lymph flow rate and the height of the outflow end of the lymph cannula. From this relationship, determined at the end of the period of elevated left atrial pressure, we calculated the effective resistance and pressure driving lymph from the lungs. We also cannulated lymph vessels in the downstream direction and estimated the effective resistance and pressure opposing flow into the part of the lymphatic system between the lung hilus and the veins (extrapulmonary lymph vessels). We found that the effective resistance of the extrapulmonary part of the lymph system (0.042 +/- 0.030 (SD) cmH2O X min X microliter-1) was large compared with the resistance of the lymph vessels from the lungs (0.026 +/- 0.027). These data indicate that the resistance of the extrapulmonary part of the lung lymph system limits the maximum flow of lymph from edematous lungs.

Outflow pressure reduces lymph flow rate from various tissues

We previously reported that the very act of cannulating a lung lymph vessel could alter the unique flow characteristics that existed within the lymphatic before cannulation. We postulated that this phenomenon could hold true for lymphatics draining any organ within the body. Since it is frequently important to know the relationship between the transmicrovascular fluid flux and true lymph flow rate, it would be critical that a cannulated lymphatic vessel have the same flow characteristics as those uncannulated vessels draining the same organ. In order to test our hypothesis we cannulated lymph vessels draining the heart, liver, small intestine, kidney, and skeletal muscle. By altering the lymphatic outflow pressure (normally related to systemic venous pressure) and by using lymphatic cannulas of various resistance, we were able to demonstrate that lymph flow varied linearly with lymphatic outflow pressure in every organ. By increasing transmicrovascular fluid flux and lymph flow rate in each organ we were also able to demonstrate that effective resistance of the lymphatic vessels and the effective pressure driving lymph flow varied as a function of the physical characteristics of the organ under investigation. Characteristic effective resistances of the heart, liver, skeletal muscle, kidney, and small intestine lymphatics decreased by 83, 40, 61, 36, and 50%, respectively. Along with these changes in effective resistance, the effective lymph driving pressure in the same organs varied by 49, 0, 257, 0, and 63%, respectively. We conclude that the lymph flow from all of the organs studied varies linearly with changes in lymphatic outflow pressure and that lymphatic resistance and lymph driving pressure vary in a unique manner in each organ depending on the physical characteristics of that organ.

Overestimation of sheep lung lymph contamination.

We have previously reported that lymph from the chronic sheep lung lymph preparation contains 25-60% lymph from nonpulmonary sources. In subsequent studies we found that the lymph flow rate from cannulated lymph vessels depends on the resistance and position of the lymph cannula. Because we did not account for these factors in our estimate of the amount of nonpulmonary lymph in the sheep lung lymph preparation, our data were not accurate. We probably overestimated the amount of nonpulmonary lymph. However, the presence of nonpulmonary lymph remains a potentially serious problem with the sheep lung lymph preparation.

High frequency ventilation and the accumulation of extravascular lung water.

INTRODUCTION Pulmonary edema, or increased lung water, adversely affects gas exchange and pulmonary compliance. Numerous safety factors exist to protect against pulmonary edema and alveolar flooding. The most important of these safety factors is thought to be the lymphatic system, which removes water and protein that filters out of the pulmonary capillaries into the interstitium of the lung [ 131. Controversy has existed with regard to how lymph is propelled through the lymphatic system and cleared from the lungs. Essentially, the question centers on whether extrinsic forces caused by movement of surrounding structures (such as muscles, arteries, heart, or lungs) propels the lymph or whether intrinsic forces (rhythmic contractions of smooth muscle in the lymph vessels) were involved. Hall et al. cannulated lymph vessels throughout the body and discovered that extrinsic forces were minor and superimposed on the major intrinsic pressure waves. This applied everywhere except in the central lymphatic system, where the pressures generated by lung expansion and contraction seemed to be the major force [4]. Respiratory movement has been felt by many to be of primary importance for pulmonary or central lymph flow [7, 11, 12, 151. However, Staub and others have felt that intrinsic forces are the most important [ 12, 141. A relatively recent ventilatory technique, high frequency ventilation (HFV), offers an opportunity to examine the importance of extrinsic respiratory tidal motion on the propulsion of lymph in the pulmonary system [2,3]. High frequency ventilation differs from conventional ventilation by using a much smaller tidal volume (often less than dead space) and higher frequencies. This technique involves minimal bulk flow of air and almost no chest wall motion. High frequency ventilation provides a way to maintain normal blood gases for extended periods of time, while maintaining a decreased or absent tidal movement. We asked the question, does marked reduction of tidal expansion and contraction of the lungs have an effect on the accumulation of EVLW in normal dogs over 48 hr?

Ether-dissolved ethiodol for lymphangiography: an experimental study

In order to decrease administration time, the viscosity of Ethiodol was reduced by mixing it with ether; this mixture was evaluated in lymphangiography of dogs. The ether-Ethiodol solution was injected into a cannulated lymph vessel of one hind limb and compared to pure Ethiodol in the other. For the ether-Ethiodol solution, the time required for complete injection was 10-30 min in most cases compared to 60-105 min for Ethiodol alone. The radiographic appearance of the lymph vessels and nodes appeared similar on both sides, and follow-up lymphangiograms in two animals after 1 and 3 weeks showed no difference from the original study. Microscopic examination of the opacified lymph nodes on both sides revealed no histologic differences. The techniques has potential clinical application.

Estimation of Lymph Flow in Extremities

The lymphatic system plays an important role in the body circulation, but our methods for studying the system in the living organism are limited, and its physiology is incompletely understood. Kinmonth 6,7 introduced lymphangiography, a method for visualization of the lymph vessels in the extremities by injection of contrast medium directly into the cannulated lymph vessel. With lymphangiography, pathological changes in the lymph vessels such as varicose lymph vessels or decrease in number of lymph vessels were demonstrated in lymphedema. The decreased capacity for flow in the lymphatic system was then explained. A method for estimation of lymph flow in the extremities would be valuable for studying this decrease, and therefore the following experimental study was performed. On the assumption that substances with the molecular size of albumin or larger are solely removed from the extravascular space by the lymphatic system, radioactive iodinated human serum albumin (RIHSA) was injected into