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We investigated the functions of tonsillar mononuclear cells (TMC) regarding whether a Haemophilus parainfluenzae (HP) outer membranes antigen (HPOM) enhances IgA-related cytokine (IFN-gamma, IL-10. and TGF-beta) production in vitro by TMC in IgA nephropathy (IgAN) patients. In addition, we examined the effect of synthetic oligodeoxynucleotide and HPOM stimulation by TMC on IgA production, whether the constant region antisense to IgA inhibits the production of IgA in vitro by TMC. Eighteen patients with IgAN and 25 patients with chronic tonsillitis (CT) from 6 to 45 years (mean age of 20.9 years) participated in this study. TMC were obtained from resected tonsils, and total and HP-specific IgA levels, along with the concentration of TGF-beta, IL-10 and IFN-gamma in the supernatant of stimulated TMC were measured by ELISA. Isolated TMC were cultured with HPOM in the presence of 23 oligodeoxynucleotides (ODNs), and the induction of total IgA and HP-specific IgA in the supernatant was also measured using ELISA. To investigate the inhibition of IgA production, TMC were cultured with HPOM and antisence to IgA. We found that IgA-related cytokine (IFN-gamma, IL-10, and TGF-beta) production by unstimulated or stimulated TMC was higher in IgAN patients than CT patients. Two types of synthetic oligodeoxynucleotides produced higher HP-specific IgA than with HPOM stimulation alone. HPOM and antisence IgA inhibited the production of total IgA and HP-specific IgA in dose-depend manner. In conclusion, IFN-gamma, TGF-beta, and IL-10 influence each other in the pathogenesis of IgAN, and infection by not only HP but other bacteria or viruses which possess specific DNA sequences such as CpG motifs induce the production of HP-specific IgA by TMC.

Human immunoglobulin A1 (IgA1), which is the predominant subtype to be deposited in glomeruli in IgA nephropathy (IgAN), has a unique mucine-like structure in its hinge region. Namely, it contains O-glycans and proline-rich peptides We previously observed underglycosylation of the hinge region in serum and deposited IgA1 in IgAN. On the other hand, clinical development and exacerbation of IgAN are frequently preceded by episodes of upper respiratory tract infection, and palatine tonsils represent the predominant immunocompetent tissue of the upper respiratory tract. Therefore, we hypothesized that tonsils were one of the origins of glomerular IgA1 in IgAN, and investigated the O-glycan structure of IgA1 produced by tonsillar lymphocytes (tonsillar IgA1). A significant increase in asialo-agalacto type O-glycans was found in the tonsillar IgA1 hinge in IgAN. These results suggest that the tonsils produce underglycosylated IgA1 molecules, which enter the bloodstream and are then deposited in the glomeruli.

High (HD) and low (LD) density cells were separated on 25% BSA gradient from tonsils of 3-6 years old children. Early B lymphocyte markers and sIg-s were found on the surface of 59-82% of the LD cells. This cell population was 5-6 times more active in DNA synthesis (3H-thymidine incorporation, DNA polymerase activity) than the HD cells. The total uptake of 3H-deoxycytidine was about the same as that of 3H-thymidine. As long as practically all thymidine taken up by the cells was immediately incorporated into DNA (90-95%), only 10-15% of deoxycytidine was incorporated into DNA under the same conditions, indicating different pool sizes for the DNA precursors. The majority of deoxycytidine (70%) was converted and incorporated as dTMP. A considerable part of labeled deoxycytidine could be detected in the soluble pool in form of nucleotides (3-8%), and in an unknown form, called substance X (8-14%). Substance X was purified by TL chromatography and identified by HPLC as deoxycytidine containing liponucleotides, probable precursors for plasmamembranes. The preferential utilisation of deoxycytidine for DNA and membrane synthesis in immature B lymphocytes draws the attention to its function in early events of lymphocyte maturation.

In order to clarify the relationship between the tonsils and pustulosis palmaris et plantaris (PPP), immunological investigations were performed focusing on keratin and antikeratin antibody. As materials, the tonsils, plantar skins and peripheral blood from the patients with PPP undergoing tonsillectomy were used. The results were summarized as follows: 1) Common keratin polypeptides reacting with monoclonal anti-keratin antibody were found both in the tonsillar epithelium and in the plantar skin. 2) High molecular weight keratin polypeptides were predominant in the tonsillar epithelium of the patients with PPP as in the plantar skin. 3) The antikeratin antibody titers in the sera of the patients with PPP were on a high level, and tended to decrease after tonsillectomy. 4) The antibody in the sera of the patients with PPP reacted with 67, 63 and 56 kd bands of keratin polypeptides. 5) The antikeratin antibody titers in the culture medium of the patients with PPP, especially in the tonsillar lymphocyte cultures, were on a high level. These results suggest the possibility that the keratin of the tonsillar epithelium and the antikeratin antibody in the serum may play an important role for pustule formation.

Glycoproteins in palatine tonsillar epithelium were examined histochemically using seven different kinds of lectins to elucidate their functional morphology. UEA-I and WGA affinities, which might be altered due to an inflammatory reaction of the stroma, were detected selectively in the epithelial cell surface. Reacting products of SBA and GS-II were observed characteristically in the spinous and granular layers. No PNA, BPA and DBA activities were found in the tonsillar epithelium. These results suggest that carbohydrates on cell membranes might be changed according to the epithelial cell differentiation and some factors from the stroma.

The autonomic nervous system in the human palatine tonsil was studied systematically as follows: 1) Using the radio binding assay method, the presence of beta-adrenergic receptors and muscarinic-cholinergic receptors was demonstrated in the tonsil. 2) Using high performance liquid chromatography, the quantity of catecholamines in the tonsillar tissue was determined for the first time. 3) The distributions of sympathetic and parasympathetic nerves in the tonsillar tissue were demonstrated by fluorescence- or enzyme-histochemistry. 4) Cases of habitual tonsillitis were compared with cases of simple tonsillitis concerning the quantities of beta-adrenergic receptors and catecholamines. As a result, the difference in quantity of beta-adrenergic receptors between the two was insignificant, but the quantity of catecholamine was greater in the former.

The N-acetyl-L-tyrosine ethyl ester (ATEe)-hydrolysing activities of tonsillar extracts in cases of recurrent acute tonsillitis decreased more than those in less severe cases of chronic tonsillitis and in cases of obstructive enlargement of the palatine tonsil. Since ATEe is hydrolysed by C1-esterase, the ATEe hydrolytic activity of the tonsil might be related to the C1-activating system of the tonsil. This indicates that tonsils in which ATEe hydrolytic activities are low, repeatedly and severely, become infected, possibly due to easy invasion of micro-organisms.

No significant differences were found between hyperplastic tonsillitis and recurrent tonsillitis in the intensity of cell-mediated immune response of tonsillar T cells, the ratio of T cell subsets, the ratio of Tac-positive cells, and the stimulation index determined after incubation in the presence of TCGF alone. However, the proportion of the cells in phase S+G2+M, the ratio of OKT9-positive cells to total tonsillar lymphocytes, and the amount of polyamines (especially putrescine) were greater in hypertrophied tonsils than in the tonsils with recurrent tonsillitis. Some characteristics of recurrent tonsillitis are discussed from these differences. Comparison of tonsillar lymphocytes from children vs. adults produced similar results. The proportion of cells in phase S+G2+M, the ratio of OKT9, and the amount of polyamines were greater in tonsils of children. In addition, differences in the function of tonsillar and peripheral lymphocytes from the patient are also discussed.

The human tonsils are responsible for the final antigen-induced differentiation of B-cells to plasma cells. Their main function could be the expansion of memory IgA-B-cells, allowing J-chain expression and final differentiation under secondary signals. IgA/J+-cells have been shown to migrate into mucosal membranes, allowing a protection against microorganisms. Recurrent tonsillitis obviously interferes with this complex immunological network, possibly resulting in a reduced mucosal protection by IgA. Furthermore, bacterial antigens can initiate the liberation of biochemical inflammatory mediators. Among these, arachidonic acid products can play a key role. They can induce epithelial damage and decreased ciliary function. Furthermore, they influence tonsillar immunological reactions.

Participation of the autonomic nerve in tonsillar focal infection was investigated by measuring neurotransmitters, receptors and microvibration. In focal infection patients, the volume of norepinephrine in the tonsil increased significantly and the number of a-adrenergic receptors decreased. These findings suggest that the focally infected tonsil exhibits a high degree of sympathetic nerve activity. The ratio of N-type in microvibration decreased systematically, which indicates some imbalance or immaturity of the autonomic nervous system in focal infection patients. Based on this data, the production mechanism of tonsillar focal infection was speculated from the point of view of the autonomic nerve.

By the immunohistochemical method using anti-keratin antibodies (KL1-antigen, 56 KD keratin; PKK1-antigen, 40-52.5 KD keratin) and electron microscopy, the changes in cell shapes and the cytokeratin components of the epithelial cells during the infiltration of lymphocytes were studied. In the surface epithelium, PKK1 reacts with only the keratinocytes in the basal layer, while KL1 stains in the spinous layer. In the neck portion of the crypt, transformed keratinocytes scattered in the spinous layer react intensely with PKK1. These cells issue prolonged cytoplasmic processes which surround the cavities filled with infiltrating lymphocytes. In the deep portion of the crypt, PKK1-positive cells interconnect with one another constituting the network of the star-shaped reticular cells. The enlarged intercellular spaces of the reticular cells are filled with many lymphocytes. Our observations suggest that the infiltration of lymphocytes into the crypt epithelium induces the change in keratin expression of the epithelial cells in the human palatine tonsils, and the change of keratin molecules makes the stratified flattened epithelium to transform into the network of star-shaped reticular cells.