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Salt and pepper to taste. (Note: this soup freezes well.) Matzoh balls were prepared according to the recipe on the back of the box of matzoh meal (Manischewitz; Jersey City, NJ).

Three separate preparations of soup were made. The completed soup was collected from all three. In addition, in order to determine at which stage the soup acquired activity, 19 samples were collected during the preparation of one batch (Table 1 ). As the mixture was inhomogeneous, several samples were collected at the same time from different regions of the pot. All samples were frozen in small aliquots and stored (−80°C) until assay.
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Table 1.

Sample Descriptions

In order to determine whether particulates accounted for the activity of the soup, attempts were made to obtain a clarified preparation. Soup that was prepared by this method could not be passed through a 0.22-μm filter. In order to remove particulates, 1-mL aliquots were centrifuged (12,000g for 15 min) in Eppendorf tubes. This resulted in a visible pellet and a clarified transparent yellow supernatant, which was aspirated for subsequent assay.

To determine which components of the soup contained inhibitor activity, samples of chicken (a leg) and a portion of each of the vegetables were boiled for approximately 1 h. The supernatant broths then were harvested, frozen, and saved for assay.

For comparison purposes, commercially available soups were obtained from a local supermarket and prepared according to the directions on the packaging. No strict quality control was performed, although each preparation was evaluated by taste and was felt to be satisfactory (if variably so).
Neutrophil Chemotaxis

Peripheral blood was collected from healthy nonsmoking volunteers under a protocol approved by the University of Nebraska Institutional Review Board and by sedimentation through dextran, as described previously.22 Neutrophils then were rinsed, suspended at 106 cells/mL in Hank’s balanced salt solution (HBSS), and used as targets for chemotaxis. Chemotaxis was performed by the modified blindwell technique using 48-well multichambers and 3-μm pore size polycarbonate filters (Nucleoprobe; Cabin John, MD), as described previously.23
Soup Inhibition of Neutrophil Chemotaxis

In order to determine whether the soup could inhibit chemotaxis, dilutions of soup (1:100) were added to the top and bottom wells of the chemotaxis chamber. Zymosan-activated serum (ZAS)24 was used as the positive chemoattractant. As a control to determine whether chicken soup had chemotactic activity, chicken soup was added directly to the bottom of the chemotaxis chamber without other chemoattractants.
Concentration Dependence of Chicken Soup Effect

Serial dilutions of chicken soup were added to neutrophils in the upper portion of the chemotaxis chamber, and ZAS (1:4) or fMet-Leu-Phe (fMLP), 10−7 mol/L (Sigma; St. Louis, MO), were used as chemoattractants.
Viability

In order to determine whether soup and its components were cytotoxic, neutrophils were prepared as if for chemotaxis and were suspended in HBSS with a 1:100 dilution of soup, component vegetables, or chicken extracts. After a 30-min incubation at 37°C, cells were collected and viability was assessed by trypan blue dye exclusion visually.
Statistics

For data sets with multiple comparisons, analysis of variance was first used to determine whether any group was significantly different, following which Student’s t test was used for comparisons that appeared to be different. Data presented are mean± SEM.
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Results

Chicken soup was found to inhibit neutrophil chemotaxis. When the completed soup (without added salt and pepper) was added to neutrophils above the membrane, to the ZAS below the membrane, or to both sides of the chemotaxis membrane, neutrophil migration to ZAS was inhibited. The effect of the chicken soup was much more marked when the diluted chicken soup was added directly to the neutrophils (Fig 1 ). The diluted chicken soup by itself had a minimal effect in stimulating neutrophil chemotaxis above background in the absence of a chemoattractant (Fig 1).
Figure 1.
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Figure 1.

Inhibition of neutrophil chemotaxis by chicken soup. ZAS (dilution, 1:4) and chicken soup (cs) (prep 1, fraction 18 diluted 1:100) were added to the top and bottom of the chemotaxis chamber in various combinations as indicated. Neutrophils were added to the top of the chamber, and neutrophil chemotaxis performed. Vertical axis: migrated neutrophils (cells per high-power field [hpf]). Horizontal axis: condition. * = p < 0.05.

The inhibitory effect of chicken soup was concentration-dependent and was observed when both ZAS and fMLP were used as chemoattractants (Fig 2 ). Interestingly, chicken soup added to the neutrophils at dilutions of> 1:20 caused a slight but significant (p < 0.05) increase in neutrophil migration toward media controls (Fig 2). Three preparations of chicken soup inhibited chemotaxis to ZAS similarly (Fig 3 ). It was not possible to remove the particulate matter from the chicken soup by filtration. In order to determine whether the solid component of the chicken soup might be responsible for the inhibition of chemotaxis, therefore, the chicken soup was clarified by centrifugation at high speed. Although there was some loss of activity, the clarified supernatants of the three chicken soup preparations retained the majority of inhibitory activity (Fig 3).
Figure 2.
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Figure 2.

Inhibition of neutrophil chemotaxis and concentration dependence. Chicken soup (preparation No. 1, fraction No. 18) was placed in varying dilutions together with neutrophils in the upper portion of the chemotaxis chamber. ZAS, fMLP, or Dulbecco’s modified Eagle’s medium (DMEM) was placed in the lower portion of the chemotaxis chamber as a chemoattractant. Chemotaxis then was performed.* = p < 0.05 compared to the highest dilution

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