Intralymphatic Immunotherapy of Pollen-Induced Rhinoconjunctivitis

A Double-Blind Placebo-Controlled Trial

Terese Hylander; Olivia Larsson; Ulla Petersson-Westin; Mia Eriksson; Susanna Kumlien Georén; Ola Winqvist; Lars-Olaf Cardell


Respiratory Research. 2016;17(10) 

In This Article


In a recent study, we demonstrated that ILIT is a safe and effective administration route for the induction of allergen tolerance and treatment of nasal allergic inflammation.[9] In the present study, we have added a new cohort of study subjects and together they reinforce the hypothesis that ILIT is effective in reducing allergic symptoms and that it is not associated with severe adverse events. In addition, we highlight a role for increased allergen-specific IgG4 affinity in successful ILIT.

During the past few years, ILIT has come forward as a less time-consuming and more cost-effective alternative to conventional subcutaneous allergen-specific immunotherapy (SCIT) for treatment of pollen-induced rhinoconjunctivitis. It has provided promising clinical results in combination with an excellent safety profile,[6,9] likely related to the drastic reduction in amount of administered allergen. This present study demonstrates that, as compared to placebo, active ILIT treatment results in improvement of seasonal allergic symptoms, without any moderate or severe adverse reactions. This study further supports ILIT as a safe and effective treatment strategy for patients with pollen-induced rhinoconjunctivitis. Indeed, the 60 % success rate is comparable to the approximate 70 % success rate for SCIT.[12] Although it is evident that five out of eight placebo patients responded to treatment, it is worth noting that the number needed to treat (NNT) value for ILIT, which takes into account responders and non-responders in both active and placebo groups, is 7.5. This NNT value is much lower than those calculated for more common treatments of allergic rhinitis, such as anti-histamines.[13]

The somewhat lower success rate of ILIT as compared to SCIT may be related to the technical aspects of allergen injections. Unlike SCIT, ILIT uses complex medical equipment and involves higher technical precision, as targeted nodes are relatively small (0.5–1.5 cm). Despite the use of a fine needle and ultrasound guidance, it can on occasion be difficult to penetrate the capsule surrounding the lymph node, which may lead to inadequate administration of allergen.

Despite repeated promising results, the overall efficacy of ILIT against grass-pollen-induced allergic rhinitis has recently been questioned.[10] In a double-blind placebo-controlled trial, 15 patients receiving active ILIT against grass pollen showed immunological indications of tolerance, but no clinical improvement. As recently argued,[14,15] these results were likely due to the interval between injections. Witten et al[10] injected allergen every 2 weeks, which, unlike a 4-week interval, does not allow sufficient time for the development of non-interfering waves of antigen-specific immune responses.[16] The present study, where injections were given at 4-week intervals, again highlights the importance of correct time interval between injections.

In this study, two distinct sub-populations could be observed among the actively treated patients. Patients in the first sub-population reported a clear improvement of their allergic symptoms, as characterised by reduced nasal allergen reactivity, decreased usage of allergy medication (anti-histamines, local steroids, eye drops) and self-recorded improvement (VAS-scale). In contrast, patients in the second sub-population experienced no resolution of symptoms. The reason for the emergence of two sub-populations is unknown and may, at least in part, be related to technical difficulties during intralymphatic injection, resulting in extra-nodal deposits of allergen and an ineffective tolerance induction. Nevertheless, the sub-populations firstly demonstrate the usefulness of the nasal provocation test (NPT) as a read-out of clinical efficacy, as nine out of 12 patients with self-reported improvement in season allergic symptoms (VAS-scale) also reported a reduction in symptoms following NPT. Secondly, these populations give insights into the underlying immunological mechanisms associated with efficacious ILIT.

The present study, as well as the aforementioned study by Witten et al,[10] both suggest that immunological alterations associated with tolerance induction, including elevated T-regulatory cells and cytokines, and IgG4 levels in blood, are not prerequisites for clinical efficacy of ILIT. Witten et al[10] noted increases in T-regulatory cells, IL-10 production and IgG4 following active ILIT, but demonstrated no clinical improvement when compared to placebo. In contrast, we demonstrated clinical improvement in the absence of increases in circulating T-regulatory cells and total serum IgG4. The reason for this is unclear, but may be related to the short time interval between vaccination and blood sampling. However, we demonstrated that clinical improvement was associated and significantly correlated with increased allergen-specific IgG4 affinity. Indeed, as previously suggested[14] affinity of B- and T-cell receptors is likely a very important parameter for the induction of tolerance to allergen and subsequent efficacious therapeutic responses.

There is an intricate interaction between B- and T-cells in the specialised lymph node compartment. B-cells bind the injected allergen via the cell-surface B-cell receptor (BCR), which leads to receptor mediated endocytosis, processing and peptide loading. Subsequently, allergen-derived peptides are presented via the MHC class II pocket to allergen-specific CD4+ T-helper cells. This B-cell induced activation of T-cells, reciprocally, allows for T-helper cell-licensed class switching and affinity maturation of IgG.[17] In the case of the present study, the injected composition resulted in increased affinity of IgG4, avoiding IgE production. The reason for IgG4 preferences in the present study is likely the use of adjuvant, as well as the compartment for the delivery of allergens, lymph nodes being more prone to IgG responses, as compared to airway responses to pollen allergens.

Furthermore, competition for antigen, which occurs more effectively when low concentrations of antigen is present in lymph follicles, is vital for the positive selection of high affinity B-cells, and this phenomenon has similarly been suggested for selection of high affinity T-cells.[18] This suggests that a 2-week-interval between injections, where concentrations of allergen in lymph nodes is consistently high, may induce clear B- and T-cell responses, as seen by increased serum levels of total IgG4 levels and allergen-induced intracellular T-cell cytokine production,[10] but with a low affinity maturation, resulting in poor clinical efficacy. Indeed, previous studies have noted that functionality of IgG4, rather than levels, more closely relate to clinical efficacy in AIT[19,20]

Although this study clearly indicated that ILIT does improve clinical symptoms of pollen-induced rhinoconjunctivitis, particularly in the improved sub-population, there are a number of limitations. Firstly, the use of a VAS scale for patient recorded outcome has been debated, in particular in long-term studies, due to recall bias. However, several publications have demonstrated that using VAS to measure self-recorded outcome following immunotherapy parallels the more objective, and more regularly monitored, symptom and medication score (SMS), up to five years following treatment.[21–23] In addition, with the small sample size of 35 patients used in this study, it is impossible determine whether ILIT is an effective treatment for the general allergic population. However, previous studies with smaller[9] and larger[6] sample sizes show similar effects, which suggests that ILIT has a generalised efficacy. Further, as this study only spanned over one year, long-term efficacy of ILIT is unknown. Nevertheless, previous studies have demonstrated that efficacy of ILIT against grass-pollen-induced rhinoconjunctivitis persists for at least 3 years following treatment.[6]